Galactopyranosyl-cyclohexyl derivatives as e-selectin antagonists

ABSTRACT

Compounds, compositions, and methods for treatment and/or prevention of at least one disease, disorder, and/or condition by inhibiting binding of an E-selectin to an E-selectin ligand are disclosed. For example, E-selectin antagonists are described and pharmaceutical compositions comprising at least one of the same.

This application claims the benefit under 35 U.S.C. § 119(e) to U.S.Provisional Application No. 62/471,860 filed Mar. 15, 2017, whichapplication is incorporated by reference herein in its entirety.

Compounds, compositions, and methods for treating and/or preventing atleast one disease, disorder, and/or condition associated with E-selectinactivity including, for example, inflammatory diseases and cancers, aredisclosed herein.

When a tissue is infected or damaged, the inflammatory process directsleukocytes and other immune system components to the site of infectionor injury. Within this process, leukocytes play an important role in theengulfment and digestion of microorganisms. The recruitment ofleukocytes to infected or damaged tissue is critical for mounting aneffective immune defense.

Selectins are a group of structurally similar cell surface receptorsimportant for mediating leukocyte binding to endothelial cells. Theseproteins are type 1 membrane proteins and are composed of an aminoterminal lectin domain, an epidermal growth factor (EGF)-like domain, avariable number of complement receptor related repeats, a hydrophobicdomain spanning region and a cytoplasmic domain. The bindinginteractions appear to be mediated by contact of the lectin domain ofthe selectins and various carbohydrate ligands.

There are three known selectins: E-selectin, P-selectin, and L-selectin.E-selectin is found on the surface of activated endothelial cells, whichline the interior wall of capillaries. E-selectin binds to thecarbohydrate sialyl-Lewis^(x) (sLe^(x)), which is presented as aglycoprotein or glycolipid on the surface of certain leukocytes(monocytes and neutrophils) and helps these cells adhere to capillarywalls in areas where surrounding tissue is infected or damaged; andE-selectin also binds to sialyl-Lewis^(a) (sLe^(a)), which is expressedon many tumor cells. P-selectin is expressed on inflamed endothelium andplatelets, and also recognizes sLe^(x) and sLe^(a), but also contains asecond site that interacts with sulfated tyrosine. The expression ofE-selectin and P-selectin is generally increased when the tissueadjacent to a capillary is infected or damaged. L-selectin is expressedon leukocytes. Selectin-mediated intercellular adhesion is an example ofa selectin-mediated function.

Although selectin-mediated cell adhesion is required for fightinginfection and destroying foreign material, there are situations in whichsuch cell adhesion is undesirable or excessive, resulting in tissuedamage instead of repair. For example, many pathologies (such asautoimmune and inflammatory diseases, shock and reperfusion injuries)involve abnormal adhesion of white blood cells. Such abnormal celladhesion may also play a role in transplant and graft rejection. Inaddition, some circulating cancer cells appear to take advantage of theinflammatory mechanism to bind to activated endothelium and metastasize.In such circumstances, modulation of selectin-mediated intercellularadhesion may be desirable.

Accordingly, there is a need in the art for inhibitors ofselectin-mediated function, function, e.g., of selectin-dependent celladhesion, and for the development of methods employing such compounds.The present disclosure may fulfill one or more of these needs and/or mayprovide other advantages.

Compounds, compositions, and methods for treating and/or preventing(i.e., reducing the likelihood of occurrence or reoccurrence) at leastone disease, disorder, and/or condition in which inhibiting binding ofE-selectin to one or more ligands may play a role are disclosed.

Disclosed are glycomimetic E-selectin antagonists of Formula (I):

prodrugs of Formula (I), and pharmaceutically acceptable salts of any ofthe foregoing, wherein R¹, R², R³, R⁴, R⁵, R⁶, L, and M are definedherein.

As used herein, ‘compound of Formula (I)’ includes E-selectinantagonists of Formula (I), pharmaceutically acceptable salts ofE-selectin antagonists of Formula (I), prodrugs of E-selectinantagonists of Formula (I), and pharmaceutically acceptable salts ofprodrugs of E-selectin antagonists of Formula (I).

In some embodiments, pharmaceutical compositions comprising at least onecompound of Formula (I) and optionally at least one additionalpharmaceutically acceptable ingredient are presented.

In some embodiments, a method for treatment and/or prevention of atleast one disease, disorder, and/or condition where inhibition ofE-selectin mediated functions is useful is disclosed, the methodcomprising administering to a subject in need thereof an effectiveamount of at least one compound of Formula (I) and/or a pharmaceuticalcomposition comprising at least one compound of Formula (I).

In the following description, certain specific details are set forth inorder to provide a thorough understanding of various embodiments.However, one skilled in the art will understand that the disclosedembodiments may be practiced without these details. In other instances,well-known structures have not been shown or described in detail toavoid unnecessarily obscuring descriptions of the embodiments. These andother embodiments will become apparent upon reference to the followingdetailed description and attached drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagram illustrating the synthesis of building block 3.

FIG. 2 is a diagram illustrating the synthesis of building block 7.

FIG. 3 is a diagram illustrating the synthesis of intermediate 11.

FIG. 4 is a diagram illustrating the synthesis of intermediate 12.

FIG. 5 is a diagram illustrating the synthesis of compound 17.

FIG. 6 is a diagram illustrating the synthesis of intermediate 16.

FIG. 7 is a diagram illustrating the synthesis of compound 24.

FIG. 8 is a diagram illustrating the synthesis of building block 27.

FIG. 9 is a diagram illustrating the synthesis of compound 30.

FIG. 10 is a diagram illustrating the synthesis of compound 33.

FIG. 11 is a diagram illustrating the synthesis of compound 35.

FIG. 12 is a diagram illustrating the synthesis of intermediate 40.

FIG. 13 is a diagram illustrating the synthesis of compound 43.

FIG. 14 is a diagram illustrating the synthesis of compound 46.

FIG. 15 is a diagram illustrating a prophetic synthesis of intermediate69.

FIG. 16 is a diagram illustrating a prophetic synthesis of compound 73.

Disclosed herein are glycomimetic E-selectin antagonists, pharmaceuticalcompositions comprising the same, and methods for inhibiting E-selectinmediated functions using the same. The compounds and compositions of thepresent disclosure may be useful for treating and/or preventing at leastone disease, disorder, and/or condition that is treatable by inhibitingbinding of E-selectin to one or more ligands.

The compounds of the present disclosure may have at least one improvedphysicochemical, pharmacological, and/or pharmacokinetic property.

In some embodiments, presented are glycomimetic E-selectin antagonistsof Formula (I):

prodrugs of Formula (I), and pharmaceutically acceptable salts of any ofthe foregoing, wherein

R¹ is chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₈haloalkyl, C₂₋₈ haloalkenyl, C₂₋₈ haloalkynyl,

groups, wherein n is chosen from integers ranging from 0 to 2, R⁷ ischosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆cycloalkylalkyl, and —C(═O)R⁸ groups, and each R⁸ is independentlychosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆cycloalkylalkyl, C₆-18 aryl, and C₁₋₁₃ heteroaryl groups;

R² is chosen from C₁₋₁₂ alkyl groups substituted with at least onesubstituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groupsand C₂₋₁₂ alkenyl groups substituted with at least one substituentchosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups, wherein eachQ is independently chosen from H and pharmaceutically acceptablecations;

R³ is chosen from —OH, —OY¹, halo, —NH₂, —NHY¹, —NY¹Y², —OC(═O)Y¹,—NHC(═O)Y¹, and —NHC(═O)NHY¹ groups, wherein Y¹ and Y², which may be thesame or different, are independently chosen from C₁₋₁₂ alkyl, C₂₋₁₂alkenyl, C₂₋₁₂ alkynyl, C₄₋₁₆ cycloalkylalkyl, C₂₋₁₂ heterocyclyl, C₆₋₁₈aryl, and C₁₋₁₃ heteroaryl groups, wherein Y¹ and Y² may join togetheralong with the nitrogen atom to which they are attached to form a ring;

R⁴ is chosen from —CN, —CH₂CN, and —C(═O)Y³ groups, wherein Y³ is chosenfrom C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, —NHOH, —NHOCH₃, —NHCN, and—NZ¹Z² groups, wherein Z¹ and Z², which may be identical or different,are independently chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂alkynyl, C₁₋₁₂ haloalkyl, C₂₋₁₂ haloalkenyl, C₂₋₁₂ haloalkynyl, andC₇₋₁₂ arylalkyl groups, wherein Z¹ and Z² may join together along withthe nitrogen atom to which they are attached to form a ring;

R⁵ is chosen from H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₁₂haloalkyl, C₂₋₁₂ haloalkenyl, C₂₋₁₂ haloalkynyl, C₄₋₁₆ cycloalkylalkyl,and C₆₋₁₈ aryl groups;

R⁶ is chosen from —CN, C₁₋₁₂ alkyl, and C₁₋₁₂ haloalkyl groups;

L is chosen from chosen from a bond, C₆₋₁₈ aryl, and C₁₋₁₂ heteroarylgroups; and

M is chosen from linker groups;

with the proviso that when L is a bond and R² is chosen from —OH and—CO₂Q groups, then M is not a steroidal moiety.

In some embodiments, R¹ is chosen from H, C₁₋₁₂ alkyl, and C₁₋₁₂haloalkyl groups. In some embodiments, R¹ is chosen from H and C₁₋₈alkyl groups. In some embodiments, R¹ is H. In some embodiments, R¹ ischosen from C₁₋₆ alkyl groups. In some embodiments, R¹ is chosen fromC₁₋₄ alkyl groups. In some embodiments, R¹ is chosen from methyl andethyl. In some embodiments, R¹ is methyl. In some embodiments, R¹ isethyl.

In some embodiments, R¹ is chosen from

In some embodiments, R¹ is chosen from

In some embodiments, R⁷ is chosen from H, C₁₋₈ alkyl, and —C(═O)R⁸groups. In some embodiments, R⁷ is chosen from H and C₁₋₈ alkyl groups.In some embodiments, R⁷ is chosen from C₁₋₄ alkyl groups. In someembodiments, R⁷ is H.

In some embodiments, each R⁸ is independently chosen from H, C₁₋₈ alkyl,C₆₋₁₈ aryl groups, and C₁₋₁₃ heteroaryl groups. In some embodiments, atleast one R⁸ is chosen from C₁₋₈ alkyl groups. In some embodiments, atleast one R⁸ is chosen from C₁₋₄ alkyl groups. In some embodiments, atleast one R⁸ is chosen from methyl and ethyl. In some embodiments, atleast one R⁸ is H. In some embodiments, at least one R⁷ is methyl. Insome embodiments, at least one R⁸ is ethyl.

In some embodiments, at least one R⁸ is chosen from

In some embodiments, R¹ is

In some embodiments, R¹ is

In some embodiments, R¹ is

In some embodiments, R² is chosen from C₁₋₁₂ alkyl groups substitutedwith at least one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q,and —SO₃Q groups. In some embodiments R² is chosen from C₂₋₁₂ alkenylgroups substituted with at least one substituent chosen from —OH,—OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments, R² ischosen from C₁₋₈ alkyl groups substituted with at least one substituentchosen —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In someembodiments R² is chosen from C₂₋₈ alkenyl groups substituted with atleast one substituent chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Qgroups. In some embodiments, R² is chosen from C₁₋₅ alkyl groupssubstituted with at least one substituent chosen from —OH, —OSO₃Q,—OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen fromC₂₋₅ alkenyl groups substituted with at least one substituent chosenfrom —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments,the at least one substituent of R² is —OH. In some embodiments, the atleast one substituent of R² is chosen from —OSO₃Q groups. In someembodiments, the at least one substituent of R² is chosen from —OPO₃Q₂groups. In some embodiments, the at least one substituent of R² ischosen from —CO₂Q groups. In some embodiments, the at least onesubstituent of R² is chosen from —SO₃Q groups.

In some embodiments, R² is chosen from C₁₋₈ alkyl groups substitutedwith at least two substituents independently chosen from —OH, —OSO₃Q,—OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen fromC₂₋₈ alkenyl groups substituted with at least two substituentsindependently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups.In some embodiments, R² is chosen from C₁₋₅ alkyl groups substitutedwith at least two substituents independently chosen from —OH, —OSO₃Q,—OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen fromC₂₋₅ alkenyl groups substituted with at least two substituentsindependently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups.In some embodiments, the at least two substituents of R² are —OH. Insome embodiments, the at least two substituents of R² are independentlychosen from —OSO₃Q groups. In some embodiments, the at least twosubstituents of R² are independent chosen from —OPO₃Q₂ groups. In someembodiments, the at least two substituents of R² are independentlychosen from —CO₂Q groups. In some embodiments, the at least twosubstituents of R² are independently chosen from —SO₃Q groups.

In some embodiments, R² is chosen from C₁₋₈ alkyl groups substitutedwith at least three substituents independently chosen from —OH, —OSO₃Q,—OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen fromC₂₋₈ alkenyl groups substituted with at least three substituentsindependently chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Q groups.In some embodiments, R² is chosen from C₁₋₅ alkyl groups substitutedwith at least three substituents independently chosen from —OH, —OSO₃Q,—OPO₃Q₂, —CO₂Q, and —SO₃Q groups. In some embodiments R² is chosen fromC₂₋₅ alkenyl groups substituted with at least three substituentsindependentaly chosen from —OH, —OSO₃Q, —OPO₃Q₂, —CO₂Q, and —SO₃Qgroups. In some embodiments, the at least three substituents of R² are—OH. In some embodiments, the at least three substituents of R² areindependently chosen from —OSO₃Q groups. In some embodiments, the atleast three substituents of R² are independently chosen from —OPO₃Q₂groups. In some embodiments, the at least three substituents of R² areindependently chosen from —CO₂Q groups. In some embodiments, the atleast three substituents of R² are independently chosen from —SO₃Qgroups.

In some embodiments, at least one Q is H. In some embodiments, at leastone Q is chosen from pharmaceutically acceptable cations. In someembodiments, at least one Q is chosen from sodium, potassium, lithium,ammonium (substituted and unsubstituted), calcium, magnesium, iron,zinc, copper, manganese, and aluminum cations. In some embodiments, atleast one Q is a sodium cation. In some embodiments, at least one Q is apotassium cation. In some embodiments, at least one Q is chosen fromammonium cations.

In some embodiments, each Q is H. In some embodiments, each Q is chosenfrom pharmaceutically acceptable cations. In some embodiments, each Q ischosen from sodium, potassium, lithium, ammonium (substituted andunsubstituted), calcium, magnesium, iron, zinc, copper, manganese, andaluminum cations. In some embodiments, each Q is a sodium cation. Insome embodiments, each Q is a potassium cation. In some embodiments,each Q is chosen from ammonium cations.

In some embodiments, R³ is chosen from —OH, —OY¹, —OC(═O)Y′, —NH₂, and—NHC(═O)Y¹ groups, wherein Y¹ is chosen from C₁₋₈ alkyl, C₄₋₁₂cycloalkylalkyl, C₂₋₈ heterocyclyl, C₆₋₁₂ aryl, and C₁₋₉ heteroarylgroups. In some embodiments, R³ is chosen from —OY¹ groups. In someembodiments, R³ is chosen from —OC(═O)Y′ groups. In some embodiments, R³is chosen from —NHC(═O)Y¹ groups. In some embodiments, R³ is —OH. Insome embodiments, R³ is —NH₂.

In some embodiments, R³ is chosen from

In some embodiments, R³ is

In some embodiments, R³ is

In some embodiments, R⁴ is —CN. In some embodiments, R⁴ is —CH₂CN. Insome embodiments, R⁴ is chosen from —C(═O)Y³ groups, wherein Y³ ischosen from —OZ¹, —NHOH, —NHOCH₃, and —NZ¹Z² groups. In someembodiments, R⁴ is chosen from —C(═O)OZ¹ groups. In some embodiments, R⁴is chosen from —C(═O)NZ¹Z² groups. In some embodiments, Z¹ and Z², whichmay be identical or different, are independently chosen from H, C₁₋₈alkyl, C₁₋₈ haloalkyl, and C₇₋₁₂ arylalkyl groups. In some embodiments,at least one of Z¹ and Z² is H. In some embodiments, each of Z¹ and Z²is H. In some embodiments, at least one of Z¹ and Z² is methyl. In someembodiments, each of Z¹ and Z² is methyl. In some embodiments, at leastone of Z¹ and Z² is ethyl. In some embodiments, each of Z¹ and Z² isethyl. In some embodiments, Z¹ is H and Z² is methyl. In someembodiments, Z¹ and Z² join together along with the nitrogen atom towhich they are attached to form a ring.

In some embodiments, R⁴ is chosen from

In some embodiments, R⁴ is chosen from

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁵ is chosen from C₁₋₁₂ alkyl groups. In someembodiments, R⁵ is chosen from C₁₋₈ alkyl groups. In some embodiments,R⁵ is chosen from C₁₋₁₂ haloalkyl groups. In some embodiments, R⁵ ischosen from C₁₋₈ haloalkyl groups. In some embodiments, R⁵ is chosenfrom C₄₋₁₆ cycloalkylalkyl groups. In some embodiments, R⁵ is chosenfrom C₄₋₈ cycloalkylalkyl groups. In some embodiments, R⁵ is chosen frompropyl, cyclopropylmethyl, and cyclohexylmethyl. In some embodiments, R⁵is propyl. In some embodiments, R⁵ is cyclopropylmethyl. In someembodiments, R⁵ is cyclohexylmethyl.

In some embodiments, R⁶ is chosen from C₁₋₁₂ alkyl groups. In someembodiments, R⁶ is chosen from C₁₋₄ alkyl groups. In some embodiments,R⁶ is chosen from C₁₋₁₂ haloalkyl groups. In some embodiments, R⁶ ischosen from C₁₋₄ haloalkyl groups. In some embodiments, R⁶ is chosenfrom halomethyl groups. In some embodiments, R⁶ is CF₃. In someembodiments, R⁶ is CH₃. In some embodiments, R⁶ is CN.

In some embodiments, L is a bond. In some embodiments L is chosen fromC₆₋₁₈ aryl groups. In some embodiments L is chosen from C₁₋₁₂ heteroarylgroups.

In some embodiments, L is

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In some embodiments, linker groups (M) may be chosen from groupscomprising spacer groups, such spacer groups as, for example,—(CH₂)_(t)— and —O(CH₂)_(t)—, wherein t is chosen from integers rangingfrom 1 to 20. Other non-limiting examples of spacer groups includecarbonyl groups and carbonyl-containing groups such as, for example,amide groups. A non-limiting example of a spacer group is:

In some embodiments, the linker group (M) is chosen from:

In some embodiments, the linker group (M) is chosen from polyethyleneglycols (PEGS), —C(═O)NH(CH₂)_(v)O—, —C(═O)NH(CH₂)_(v)NHC(═O)—,—C(═O)NHC(═O)(CH₂)NH— and —C(═O)NH(CH₂)_(v)C(═O)NH— groups, wherein v ischosen from integers ranging from 2 to 20. In some embodiments, v ischosen from integers ranging from 2 to 4. In some embodiments, v is 2.In some embodiments, v is 3. In some embodiments, v is 4.

In some embodiments, the linker group (M) is:

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Also provided are pharmaceutical compositions comprising at least onecompound of Formula (I). Such pharmaceutical compositions are describedin greater detail herein. These compounds and compositions may be usedin the methods described herein.

In some embodiments, a method for treating and/or preventing at leastone disease, disorder, and/or condition where inhibition of E-selectinmediated functions may be useful is disclosed, the method comprisingadministering at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I).

In some embodiments, a method for treating and/or preventing at leastone inflammatory disease, disorder, and/or condition in which theadhesion and/or migration of cells occurs in the disease, disorder,and/or condition is disclosed, the method comprising administering atleast one compound of Formula (I) and/or a pharmaceutical compositioncomprising at least one compound of Formula (I).

In some embodiments, a method for inhibiting adhesion of a cancer cellthat expresses a ligand of E-selectin to an endothelial cell expressingE-selectin on the cell surface of the endothelial cell is disclosed, themethod comprising contacting the endothelial cell and at least onecompound of Formula (I) and/or a pharmaceutical composition comprisingat least one compound of Formula (I) such that the at least one compoundof Formula (I) interacts with E-selectin on the endothelial cell,thereby inhibiting binding of the cancer cell to the endothelial cell.In some embodiments, the endothelial cell is present in the bone marrow.

In some embodiment, a method for treating and/or preventing a cancer isdisclosed, the method comprising administering to a subject in needthereof an effective amount of at least one compound of Formula (I)and/or a pharmaceutical composition comprising at least one compound ofFormula (I). In some embodiments, at least one compound of Formula (I)and/or pharmaceutical composition comprising at least one compound ofFormula (I) may be administered in conjunction with (i.e., as an adjuncttherapy, which is also called adjunctive therapy) chemotherapy and/orradiotherapy.

The chemotherapy and/or radiotherapy may be referred to as the primaryanti-tumor or anti-cancer therapy that is being administered to thesubject to treat the particular cancer. In some embodiments, a methodfor reducing (i.e., inhibiting, diminishing) chemosensitivity and/orradiosensitivity of hematopoietic stem cells (HSC) to thechemotherapeutic drug(s) and/or radiotherapy, respectively, isdisclosed, the method comprising administering to a subject in needthereof an effective amount of at least one compound of Formula (I)and/or a pharmaceutical composition comprising at least one compound ofFormula (I).

In some embodiments, a method for enhancing (i.e., promoting) survivalof hematopoietic stem cells is provided, the method comprisingadministering to a subject in need thereof at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I).

In some embodiments, a method for decreasing the likelihood ofoccurrence of metastasis of cancer cells (also called tumor cellsherein) in a subject who is in need thereof is disclosed, the methodcomprising administering an effective amount of at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I).

In some embodiments, a method for treatment and/or prevention of atleast one cancer in which the cancer cells may leave the primary site isdisclosed, the method comprising administering to a subject in needthereof an effective amount of at least one compound of Formula (I)and/or a pharmaceutical composition comprising at least one compound ofFormula (I). A primary site may be, for example, solid tissue (e.g.,breast or prostate) or the bloodstream.

In some embodiments, a method for treatment and/or prevention of atleast one cancer in which it is desirable to mobilize cancer cells froma site into the bloodstream and/or retain the cancer cells in thebloodstream is disclosed, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I).

In some embodiments, a method for decreasing the likelihood ofoccurrence of infiltration of cancer cells into bone marrow isdisclosed, the method comprises administering to a subject in needthereof an effective amount of at least one compound of Formula (I)and/or a pharmaceutical composition comprising at least one compound ofFormula (I).

In some embodiments, a method for releasing cells into circulating bloodand enhancing retention of the cells in the blood is disclosed, themethod comprising administering to a subject in need thereof aneffective amount of at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I). In some embodiments, the method further includes collecting thereleased cells. In some embodiments, collecting the released cellsutilizes apheresis. In some embodiments, the released cells are stemcells (e.g., bone marrow progenitor cells). In some embodiments, G-CSFis administered to the individual.

In some embodiments, a method for treating and/or preventing thrombosisis disclosed, the method comprising administering to a subject in needthereof an effective amount of at least one compound of Formula (I)and/or a pharmaceutical composition comprising at least one compound ofFormula (I).

In some embodiments, a method for treating and/or preventing mucositisis disclosed, the method comprising administering to a subject in needthereof an effective amount of at least one compound of Formula (I)and/or a pharmaceutical composition comprising at least one compound ofFormula (I).

In some embodiments, a method for treating and/or preventing onecardiovascular disease, disorder and/or condition is disclosed, themethod comprising administering to a subject in need thereof aneffective amount of at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I).

In some embodiments, a method for treatment and/or prevention ofatherosclerosis is disclosed, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I).

In some embodiments, a compound of Formula (I) and/or a pharmaceuticalcomposition comprising at least one compound of Formula (I) may be usedfor the preparation and/or manufacture of a medicament for use intreating and/or preventing at least one of the diseases, disorders,and/or conditions described herein.

Whenever a term in the specification is identified as a range (e.g.,C₁₋₄ alkyl) or “ranging from”, the range independently discloses andincludes each element of the range. As a non-limiting example, C₁₋₄alkyl groups includes, independently, C₁ alkyl groups, C₂ alkyl groups,C₃ alkyl groups, and C₄ alkyl groups. As another non-limiting example,“n is an integer ranging from 0 to 2” includes, independently, 0, 1, and2.

The term “at least one” refers to one or more, such as one, two, etc.For example, the term “at least one C₁₋₄ alkyl group” refers to one ormore C₁₋₄ alkyl groups, such as one C₁₋₄ alkyl group, two C₁₋₄ alkylgroups, etc.

The term “alkyl” includes saturated straight, branched, and cyclic (alsoidentified as cycloalkyl), primary, secondary, and tertiary hydrocarbongroups. Non-limiting examples of alkyl groups include methyl, ethyl,propyl, isopropyl, cyclopropyl, butyl, secbutyl, isobutyl, tertbutyl,cyclobutyl, 1-methylbutyl, 1,1-dimethylpropyl, pentyl, cyclopentyl,isopentyl, neopentyl, cyclopentyl, hexyl, isohexyl, and cyclohexyl.Unless stated otherwise specifically in the specification, an alkylgroup may be optionally substituted.

The term “alkenyl” includes straight, branched, and cyclic hydrocarbongroups comprising at least one double bond. The double bond of analkenyl group can be unconjugated or conjugated with another unsaturatedgroup. Non-limiting examples of alkenyl groups include vinyl, allyl,butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl,2-ethylhexenyl, and cyclopent-1-en-1-yl. Unless stated otherwisespecifically in the specification, an alkenyl group may be optionallysubstituted.

The term “alkynyl” includes straight and branched hydrocarbon groupscomprising at least one triple bond. The triple bond of an alkynyl groupcan be unconjugated or conjugated with another unsaturated group.Non-limiting examples of alkynyl groups include ethynyl, propynyl,butynyl, pentynyl, and hexynyl. Unless stated otherwise specifically inthe specification, an alkynyl group may be optionally substituted.

The term “aryl” includes hydrocarbon ring system groups comprising atleast 6 carbon atoms and at least one aromatic ring. The aryl group maybe a monocyclic, bicyclic, tricyclic or tetracyclic ring system, whichmay include fused or bridged ring systems. Non-limiting examples of arylgroups include aryl groups derived from aceanthrylene, acenaphthylene,acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene,fluorene, as-indacene, s-indacene, indane, indene, naphthalene,phenalene, phenanthrene, pleiadene, pyrene, and triphenylene. Unlessstated otherwise specifically in the specification, an aryl group may beoptionally substituted.

The terms “E-selectin antagonist” and “E-selectin inhibitor” are usedinterchangeably herein, and include inhibitors of E-selectin only, aswell as inhibitors of E-selectin and either P-selectin or L-selectin,and inhibitors of E-selectin, P-selectin, and L-selectin.

The term “glycomimetic” includes any naturally occurring ornon-naturally occurring carbohydrate compound in which at least onesubstituent has been replaced, or at least one ring has been modified(e.g., substitution of carbon for a ring oxygen), to yield a compoundthat is not fully carbohydrate.

The term “halo” or “halogen” includes fluoro, chloro, bromo, and iodo.

The term “haloalkyl” includes alkyl groups, as defined herein,substituted by at least one halogen, as defined herein. Non-limitingexamples of haloalkyl groups include trifluoromethyl, difluoromethyl,trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl,3-bromo-2-fluoropropyl, and 1,2-dibromoethyl. A “fluoroalkyl” is ahaloalkyl wherein at least one halogen is fluoro. Unless statedotherwise specifically in the specification, a haloalkyl group may beoptionally substituted.

The term “haloalkenyl” includes alkenyl groups, as defined herein,substituted by at least one halogen, as defined herein. Non-limitingexamples of haloalkenyl groups include fluoroethenyl,1,2-difluoroethenyl, 3-bromo-2-fluoropropenyl, and 1,2-dibromoethenyl. A“fluoroalkenyl” is a haloalkenyl substituted with at least one fluorogroup. Unless stated otherwise specifically in the specification, ahaloalkenyl group may be optionally substituted.

The term “haloalkynyl” includes alkynyl groups, as defined herein,substituted by at least one halogen, as defined herein. Non-limitingexamples include fluoroethynyl, 1,2-difluoroethynyl,3-bromo-2-fluoropropynyl, and 1,2-dibromoethynyl. A “fluoroalkynyl” is ahaloalkynyl wherein at least one halogen is fluoro. Unless statedotherwise specifically in the specification, a haloalkynyl group may beoptionally substituted.

The term “heterocyclyl” or “heterocyclic ring” includes 3- to24-membered saturated or partially unsaturated non-aromatic ring groupscomprising 2 to 23 ring carbon atoms and 1 to 8 ring heteroatom(s) eachindependently chosen from N, O, and S. Unless stated otherwisespecifically in the specification, the heterocyclyl groups may bemonocyclic, bicyclic, tricyclic or tetracyclic ring systems, which mayinclude fused or bridged ring systems, and may be partially or fullysaturated; any nitrogen, carbon or sulfur atom(s) in the heterocyclylgroup may be optionally oxidized; any nitrogen atom in the heterocyclylgroup may be optionally quaternized; and the heterocyclyl groupNon-limiting examples of heterocyclic ring include dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, a heterocyclyl group may be optionally substituted.

The term “heteroaryl” includes 5- to 14-membered ring groups comprising1 to 13 ring carbon atoms and 1 to 6 ring heteroatom(s) eachindependently chosen from N, O, and S, and at least one aromatic ring.Unless stated otherwise specifically in the specification, theheteroaryl group may be a monocyclic, bicyclic, tricyclic or tetracyclicring system, which may include fused or bridged ring systems; and thenitrogen, carbon or sulfur atoms in the heteroaryl radical may beoptionally oxidized; the nitrogen atom may be optionally quaternized.Non-limiting examples include azepinyl, acridinyl, benzimidazolyl,benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl,benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl,1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl,benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl,benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl,imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl,isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl,oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl,1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl,1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl,phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl,pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl,quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl,thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, andthiophenyl (i.e. thienyl). Unless stated otherwise specifically in thespecification, a heteroaryl group may be optionally substituted.

The term “pharmaceutically acceptable salts” includes both acid and baseaddition salts. Non-limiting examples of pharmaceutically acceptableacid addition salts include chlorides, bromides, sulfates, nitrates,phosphates, sulfonates, methane sulfonates, formates, tartrates,maleates, citrates, benzoates, salicylates, and ascorbates. Non-limitingexamples of pharmaceutically acceptable base addition salts includesodium, potassium, lithium, ammonium (substituted and unsubstituted),calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts.Pharmaceutically acceptable salts may, for example, be obtained usingstandard procedures well known in the field of pharmaceuticals.

The term “prodrug” includes compounds that may be converted, forexample, under physiological conditions or by solvolysis, to abiologically active compound described herein. Thus, the term “prodrug”includes metabolic precursors of compounds described herein that arepharmaceutically acceptable. A discussion of prodrugs can be found, forexample, in Higuchi, T., et al., “Pro-drugs as Novel Delivery Systems,”A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in DrugDesign, ed. Edward B. Roche, American Pharmaceutical Association andPergamon Press, 1987. The term “prodrug” also includes covalently bondedcarriers that release the active compound(s) as described herein in vivowhen such prodrug is administered to a subject. Non-limiting examples ofprodrugs include ester and amide derivatives of hydroxy, carboxy,mercapto and amino functional groups in the compounds described herein.

The term “substituted” includes the situation where, in any of the abovegroups, at least one hydrogen atom is replaced by a non-hydrogen atomsuch as, for example, a halogen atom such as F, Cl, Br, and I; an oxygenatom in groups such as hydroxyl groups, alkoxy groups, and ester groups;a sulfur atom in groups such as thiol groups, thioalkyl groups, sulfonegroups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groupssuch as amines, amides, alkylamines, dialkylamines, arylamines,alkylarylamines, diarylamines, N-oxides, imides, and enamines; a siliconatom in groups such as trialkylsilyl groups, dialkylarylsilyl groups,alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatomsin various other groups. “Substituted” also includes the situationwhere, in any of the above groups, at least one hydrogen atom isreplaced by a higher-order bond (e.g., a double- or triple-bond) to aheteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups;and nitrogen in groups such as imines, oximes, hydrazones, and nitriles.

The present disclosure includes within its scope all the possiblegeometric isomers, e.g., Z and E isomers (cis and trans isomers), of thecompounds as well as all the possible optical isomers, e.g.,diastereomers and enantiomers, of the compounds. Furthermore, thepresent disclosure includes in its scope both the individual isomers andany mixtures thereof, e.g., racemic mixtures. The individual isomers maybe obtained using the corresponding isomeric forms of the startingmaterial or they may be separated after the preparation of the endcompound according to conventional separation methods. For theseparation of optical isomers, e.g., enantiomers, from the mixturethereof conventional resolution methods, e.g., fractionalcrystallization, may be used.

The present disclosure includes within its scope all possible tautomers.Furthermore, the present disclosure includes in its scope both theindividual tautomers and any mixtures thereof.

Compounds of Formula (I) may be prepared as shown in, for example, FIGS.5, 7, 9-11, and 13-14. It is understood that one of ordinary skill inthe art may be able to make these compounds by similar methods or bycombining other methods known to one of ordinary skill in the art. It isalso understood that one of ordinary skill in the art would be able tomake other compounds of Formula (I) not specifically illustrated hereinby using appropriate starting components and modifying the parameters ofthe synthesis as needed (e.g., see FIG. 16). In general, startingcomponents may be obtained from sources such as Sigma Aldrich, AlfaAesar, Maybridge, Matrix Scientific, TCI, and Fluorochem USA, etc.and/or synthesized according to sources known to those of ordinary skillin the art (see, for example, Advanced Organic Chemistry: Reactions,Mechanisms, and Structure, 5th edition (Wiley, December 2000)) and/orprepared as described herein.

It will also be appreciated by those skilled in the art that in theprocesses described herein the functional groups of intermediatecompounds may need to be protected by suitable protecting groups, evenif not specifically described. Such functional groups include hydroxy,amino, mercapto, and carboxylic acid. Suitable protecting groups forhydroxy include but are not limited to trialkylsilyl or diarylalkylsilyl(for example, t-butyldimethylsilyl, t-butyldiphenylsilyl ortrimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitableprotecting groups for amino, amidino and guanidino include but are notlimited to t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitableprotecting groups for mercapto include but are not limited to —C(O)R″(where R″ is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl and thelike. Suitable protecting groups for carboxylic acid include but are notlimited to alkyl, aryl or arylalkyl esters. Protecting groups may beadded or removed in accordance with standard techniques, which are knownto one skilled in the art and as described herein. The use of protectinggroups is described in detail in Green, T. W. and P.G.M. Wutz,Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As one ofskill in the art would appreciate, the protecting group may also be apolymer resin such as a Wang resin, Rink resin or a2-chlorotrityl-chloride resin.

Analogous reactants to those described herein may be identified throughthe indices of known chemicals prepared by the Chemical Abstract Serviceof the American Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (the AmericanChemical Society, Washington, D.C., may be contacted for more details).Chemicals that are known but not commercially available in catalogs maybe prepared by custom chemical synthesis houses, where many of thestandard chemical supply houses (e.g., those listed above) providecustom synthesis services. A reference for the preparation and selectionof pharmaceutical salts of the present disclosure is P. H. Stahl & C. G.Wermuth “Handbook of Pharmaceutical Salts,” Verlag Helvetica ChimicaActa, Zurich, 2002.

Methods known to one of ordinary skill in the art may be identifiedthrough various reference books, articles, and databases. Suitablereference books and treatise that detail the synthesis of reactantsuseful in the preparation of compounds of the present disclosure, orprovide references to articles that describe the preparation, includefor example, “Synthetic Organic Chemistry,” John Wiley & Sons, Inc., NewYork; S. R. Sandler et al., “Organic Functional Group Preparations,” 2ndEd., Academic Press, New York, 1983; H. O. House, “Modern SyntheticReactions”, 2nd Ed., W. A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L.Gilchrist, “Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, NewYork, 1992; J. March, “Advanced Organic Chemistry: Reactions, Mechanismsand Structure,” 4th Ed., Wiley-Interscience, New York, 1992. Additionalsuitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds of the presentdisclosure, or provide references to articles that describe thepreparation, include for example, Fuhrhop, J. and Penzlin G. “OrganicSynthesis: Concepts, Methods, Starting Materials”, Second, Revised andEnlarged Edition (1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman,R. V. “Organic Chemistry, An Intermediate Text” (1996) Oxford UniversityPress, ISBN 0-19-509618-5; Larock, R. C. “Comprehensive OrganicTransformations: A Guide to Functional Group Preparations” 2nd Edition(1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. “Advanced OrganicChemistry: Reactions, Mechanisms, and Structure” 4th Edition (1992) JohnWiley & Sons, ISBN: 0-471-60180-2; Otera, J. (editor) “Modern CarbonylChemistry” (2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. “Patai's1992 Guide to the Chemistry of Functional Groups” (1992) InterscienceISBN: 0-471-93022-9; Quin, L. D. et al. “A Guide to OrganophosphorusChemistry” (2000) Wiley-Interscience, ISBN: 0-471-31824-8; Solomons, T.W. G. “Organic Chemistry” 7th Edition (2000) John Wiley & Sons, ISBN:0-471-19095-0; Stowell, J. C., “Intermediate Organic Chemistry” 2ndEdition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; “IndustrialOrganic Chemicals: Starting Materials and Intermediates: An Ullmann'sEncyclopedia” (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8volumes; “Organic Reactions” (1942-2000) John Wiley & Sons, in over 55volumes; and “Chemistry of Functional Groups” John Wiley & Sons, in 73volumes.

Biological activity of a compound described herein may be determined,for example, by performing at least one in vitro and/or in vivo studyroutinely practiced in the art and described herein or in the art. Invitro assays include without limitation binding assays, immunoassays,competitive binding assays, and cell based activity assays.

An inhibition assay may be used to screen for antagonists of E-selectin.For example, an assay may be performed to characterize the capability ofa compound described herein to inhibit (i.e., reduce, block, decrease,or prevent in a statistically or biologically significant manner)interaction of E-selectin with sLe^(a) or sLe^(x). The inhibition assaymay be a competitive binding assay, which allows the determination ofIC₅₀ values. By way of example, E-selectin/Ig chimera may be immobilizedonto a matrix (e.g., a multi-well plate, which may be made from apolymer, such as polystyrene; a test tube, and the like); a compositionmay be added to reduce nonspecific binding (e.g., a compositioncomprising non-fat dried milk or bovine serum albumin or other blockingbuffer routinely used by a person skilled in the art); the immobilizedE-selectin may be contacted with the candidate compound in the presenceof sLe^(a) comprising a reporter group under conditions and for a timesufficient to permit sLe^(a) to bind to the immobilized E-selectin; theimmobilized E-selectin may be washed; and the amount of sLe^(a) bound toimmobilized E-selectin may be detected. Variations of such steps can bereadily and routinely accomplished by a person of ordinary skill in theart.

Conditions for a particular assay include temperature, buffers(including salts, cations, media), and other components that maintainthe integrity of any cell used in the assay and the compound, which aperson of ordinary skill in the art will be familiar and/or which can bereadily determined. A person of ordinary skill in the art also readilyappreciates that appropriate controls can be designed and included whenperforming the in vitro methods and in vivo methods described herein.

The source of a compound that is characterized by at least one assay andtechniques described herein and in the art may be a biological samplethat is obtained from a subject who has been treated with the compound.The cells that may be used in the assay may also be provided in abiological sample. A “biological sample” may include a sample from asubject, and may be a blood sample (from which serum or plasma may beprepared), a biopsy specimen, one or more body fluids (e.g., lunglavage, ascites, mucosal washings, synovial fluid, urine), bone marrow,lymph nodes, tissue explant, organ culture, or any other tissue or cellpreparation from the subject or a biological source. A biological samplemay further include a tissue or cell preparation in which themorphological integrity or physical state has been disrupted, forexample, by dissection, dissociation, solubilization, fractionation,homogenization, biochemical or chemical extraction, pulverization,lyophilization, sonication, or any other means for processing a samplederived from a subject or biological source. In some embodiments, thesubject or biological source may be a human or non-human animal, aprimary cell culture (e.g., immune cells), or culture adapted cell line,including but not limited to, genetically engineered cell lines that maycontain chromosomally integrated or episomal recombinant nucleic acidsequences, immortalized or immortalizable cell lines, somatic cellhybrid cell lines, differentiated or differentiatable cell lines,transformed cell lines, and the like.

As described herein, methods for characterizing E-selectin includeanimal model studies. Non-limiting examples of animal models for liquidcancers used in the art include multiple myeloma (see, e.g., DeWeerdt,Nature 480:S38-S39 (15 Dec. 2011) doi:10.1038/480S38a; Published online14 Dec. 2011; Mitsiades et al., Clin. Cancer Res. 2009 15:1210021(2009)); acute myeloid leukemia (AML) (Zuber et al., Genes Dev. 2009Apr. 1; 23(7): 877-889). Animal models for acute lymphoblastic leukemia(ALL) have been used by persons of ordinary skill in the art for morethan two decades. Numerous exemplary animal models for solid tumorcancers are routinely used and are well known to persons of ordinaryskill in the art.

The compounds of the present disclosure and the pharmaceuticalcompositions comprising at least one of such compounds may be useful inmethods for treating and/or preventing a disease or disorder that istreatable by inhibiting at least one activity of E-selectin (and/orinhibiting binding of E-selectin to ligand(s), which in turn inhibits abiological activity).

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful in methods fortreating and/or preventing at least one inflammatory disease.Inflammation comprises reaction of vascularized living tissue to injury.By way of example, although E-selectin mediated cell adhesion may beimportant to the body's anti-infective immune response, in othercircumstances, E-selectin mediated cell adhesion may be undesirable orexcessive, resulting in tissue damage and/or scarring instead of repair.For example, many pathologies (such as autoimmune and inflammatorydiseases, shock and reperfusion injuries) involve abnormal adhesion ofwhite blood cells. Therefore, inflammation affects blood vessels andadjacent tissues in response to an injury or abnormal stimulation by aphysical, chemical, or biological agent. Examples of inflammatorydiseases, disorders, or conditions include, without limitation,dermatitis, chronic eczema, psoriasis, multiple sclerosis, rheumatoidarthritis, systemic lupus erythematosus, graft versus host disease,sepsis, diabetes, atherosclerosis, Sjogren's syndrome, progressivesystemic sclerosis, scleroderma, acute coronary syndrome, ischemicreperfusion, Crohn's disease, inflammatory bowel disease, endometriosis,glomerulonephritis, myasthenia gravis, idiopathic pulmonary fibrosis,asthma, allergic reaction, acute respiratory distress syndrome (ARDS) orother acute leukocyte-mediated lung injury, vasculitis, or inflammatoryautoimmune myositis. Other diseases and disorders for which thecompounds described herein may be useful for treating and/or preventinginclude hyperactive coronary circulation, microbial infection, cancermetastasis, thrombosis, wounds, burns, spinal cord damage, digestivetract mucous membrane disorders (e.g., gastritis, ulcers), osteoporosis,osteoarthritis, septic shock, traumatic shock, stroke, nephritis, atopicdermatitis, frostbite injury, adult dyspnoea syndrome, ulcerativecolitis, diabetes and reperfusion injury following ischemic episodes,prevention of restenosis associated with vascular stenting, and forundesirable angiogenesis, for example, angiogenesis associated withtumor growth.

As discussed in detail herein, a disease or disorder to be treated orprevented is a cancer and related metastasis and includes cancers thatcomprise solid tumor(s) and cancers that comprise liquid tumor(s). Thecompounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful in methods forpreventing and/or treating cancer. In some embodiments, the at least onecompound may be used for treating and/or preventing metastasis and/orfor inhibiting (slowing, retarding, or preventing) metastasis of cancercells.

In some embodiments, at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) is administered to a cancer patient in remission. In someembodiments, the at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) is administered as a cancer vaccine to stimulate marrow infiltratinglymphocytes (“MILs”) in a cancer patient or cancer survivor to preventrelapse.

In some embodiments, a method of treating cancer and/or preventing acancer relapse is disclosed, wherein the method comprises administeringto a patient in need thereof an effective amount of at least onecompound of Formula (I) and/or a pharmaceutical composition comprisingat least one compound of Formula (I), wherein the amount of compound ofFormula (I) administered is sufficient to mobilize MILs of the patientinto the peripheral blood.

In some embodiments, a method of treating cancer and/or preventing acancer relapse is provided comprising administering to a donor patientat least one compound of Formula (I) and/or a pharmaceutical compositioncomprising at least one compound of Formula (I) in an amount ofsufficient to mobilize MILs of the patient out of the marrow (e.g., intothe peripheral blood), recovering MILS (e.g., recovering them from theperipheral blood), and transplanting at least a portion of the MIL cellpopulation to the donor patient or another patient. In some embodiments,the MIL cell population is expanded ex vivo before transplantation.

In some embodiments, a method of preventing cancer is providedcomprising administering to a donor patient at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I) in an amount sufficient to mobilize MILs of thepatient out of the bone marrow (e.g., into the peripheral blood),recovering MILs (e.g., recovering them from the peripheral blood), andtransplanting at least a portion of MIL cell population to a subject(e.g., a non-cancer patient, a patient suffering from a different formor type of cancer than the donor patient, etc.). In some embodiments,the MIL cell population is expanded ex vivo before transplantation.

In some embodiments, the compounds of present disclosure andpharmaceutical compositions comprising at least one such compound may beused for decreasing (i.e., reducing) the likelihood of occurrence ofmetastasis of cancer cells in an individual (i.e., subject, patient) whois in need thereof. The compounds of the present disclosure andcompositions comprising at least one such compound may be used fordecreasing (i.e., reducing) the likelihood of occurrence of infiltrationof cancer cells into bone marrow in an individual who is in needthereof. The individuals (or subjects) in need of such treatmentsinclude subjects who have been diagnosed with a cancer, which includescancers that comprise solid tumor(s) and cancers that comprise liquidtumor(s).

Non-limiting examples of cancers include colorectal cancers, livercancers, gastric cancers, lung cancers, brain cancers, kidney cancers,bladder cancers, thyroid cancers, prostate cancers, ovarian cancers,cervical cancers, uterine cancers, endometrial cancers, melanomas,breast cancers, and pancreatic cancers. Liquid tumors can occur in theblood, bone marrow, the soft, sponge-like tissue in the center of mostbones, and lymph nodes and include leukemias (e.g., AML, ALL, CLL, andCML), lymphomas, and myelomas (e.g., multiple myeloma). Lymphomasinclude Hodgkin lymphoma, which is marked by the presence of a type ofcell called the Reed-Sternberg cell, and non-Hodgkin lymphomas, whichincludes a large, diverse group of cancers of immune system cells.Non-Hodgkin lymphomas can be further divided into cancers that have anindolent (slow-growing) course and those that have an aggressive(fast-growing) course, and which subtypes respond to treatmentdifferently.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be administered as an adjuncttherapy to chemotherapy and/or radiotherapy, which is/are beingdelivered to the subject as primary therapy for treating the cancer. Thechemotherapy and/or radiotherapy that may be administered depend uponseveral factors including the type of cancer, location of the tumor(s),stage of the cancer, age and gender and general health status of thesubject. A person of ordinary skill in the medical art can readilydetermine the appropriate chemotherapy regimen and/or radiotherapyregimen for the subject in need. The person of ordinary skill in themedical art can also determine, with the aid of preclinical and clinicalstudies, when the compound of the present disclosure or pharmaceuticalcomposition comprising at least one such compound should be administeredto the subject, that is whether the compound or composition isadministered prior to, concurrent with, or subsequent to a cycle of theprimary chemotherapy or radiation treatment.

Also provided herein is a method for inhibiting adhesion of a tumor cellthat expresses a ligand of E-selectin to an endothelial cell expressingE-selectin on its cell surface, which method comprises contacting theendothelial cell with at least one compound of the present disclosure orpharmaceutical compositions comprising at least one such compound,thereby permitting the compound to interact with E-selectin on theendothelial cell surface and inhibiting binding of the tumor cell to theendothelial cell. Without wishing to be bound by theory, inhibitingadhesion of tumor cells to endothelial cells may reduce in a significantmanner, the capability of the tumor cells to extravasate into otherorgans, blood vessels, lymph, or bone marrow and thereby reduce,decrease, or inhibit, or slow the progression of the cancer, includingreducing, decreasing, inhibiting, or slowing metastasis.

In some embodiments, a method for inhibiting adhesion of metastasizedtumor cells is disclosed, the method comprising administering at leastone compound of Formula (I) and/or a pharmaceutical compositioncomprising at least one compound of Formula (I).

As described herein, at least one of the compounds of the presentdisclosure or pharmaceutical compositions comprising at least one suchcompound may be administered in combination with at least one additionalanti-cancer agent. Chemotherapy may comprise one or morechemotherapeutic agents. For example, chemotherapy agents,radiotherapeutic agents, inhibitors of phosphoinositide-3 kinase (PI3K),and inhibitors of VEGF may be used in combination with a compound ofFormula (I) described herein. Non-limiting examples of inhibitors ofPI3K include the compound named by Exelixis as “XL499.” Non-limitingexamples of VEGF inhibitors include the compound called “cabo”(previously known as XL184). Many other chemotherapeutics are smallorganic molecules. As understood by a person of ordinary skill in theart, chemotherapy may also refer to a combination of two or morechemotherapeutic molecules that are administered coordinately and whichmay be referred to as combination chemotherapy. Numerouschemotherapeutic drugs are used in the oncology art and include, forexample, alkylating agents; antimetabolites; anthracyclines, plantalkaloids; and topoisomerase inhibitors.

The compounds of the present disclosure or pharmaceutical compositionscomprising at least one such compound may function independently fromthe anti-cancer agent or may function in coordination with theanti-cancer agent, e.g., by enhancing effectiveness of the anti-canceragent or vice versa. Accordingly, provided herein are methods forenhancing (i.e., enhancing, promoting, improving the likelihood of,enhancing in a statistically or biologically significant manner) and/ormaintaining survival of hematopoietic stem cells (HSC) in a subject whois treated with and/or will be treated with a chemotherapeutic drug(s)and/or radioactive therapy, respectively, comprising administering atleast one compound of Formula (I) as described herein. In someembodiments, the subject receives and/or will receive both chemotherapyand radiation therapy. Also, provided herein is a method for reducing(i.e., reducing, inhibiting, diminishing in a statistically orbiologically significant manner) chemosensitivity and/orradiosensitivity of hematopoietic stem cells (HSC) to thechemotherapeutic drug(s) and/or radioactive therapy, respectively, in asubject. Because repeated cycles of chemotherapy and radiotherapy oftendiminish the ability of HSCs to recover and replenish bone marrow, theglycomimetic compounds described herein may be useful for subjects whowill receive more than one cycle, such as at least two, three, four ormore cycles, of chemotherapy and/or radiotherapy. HSCs reside in thebone marrow and generate the cells that are needed to replenish theimmune system and the blood. Anatomically, bone marrow comprises avascular niche that is adjacent to bone endothelial sinuses (see, e.g.,Kiel et al., Cell 121:1109-21 (2005); Sugiyama et al., Immunity25:977-88 (2006); Mendez-Ferrer et al., Nature 466:829-34 (2010); Butleret al., Cell Stem Cell 6:251-64 (2010)). A recent study describes thatE-selectin promotes HSC proliferation and is an important component ofthe vascular niche (see, e.g., Winkler et al., Nature Medicine publishedonline 21 Oct. 2012; doi:10.1038/nm.2969). Deletion or inhibition ofE-selectin enhanced HSC survival in mice that were treated withchemotherapeutic agents or radiotherapy and accelerated blood neutrophilrecovery (see, e.g., Winkler et al., supra).

In addition, the administration of at least one compound of the presentdisclosure or pharmaceutical composition comprising at least one suchcompounds may be in conjunction with one or more other therapies, e.g.,for reducing toxicities of therapy. For example, at least one palliativeagent to counteract (at least in part) a side effect of a therapy (e.g.,anti-cancer therapy) may be administered. Agents (chemical orbiological) that promote recovery, or counteract side effects ofadministration of antibiotics or corticosteroids, are examples of suchpalliative agents. At least one compound described herein may beadministered before, after, or concurrently with administration of atleast one additional anti-cancer agent or at least one palliative agentto reduce a side effect of therapy. When administration is concurrent,the combination may be administered from a single container or two (ormore) separate containers.

Cancer cells (also called herein tumor cells) that may be prevented(i.e., inhibited, slowed) from metastasizing, from adhering to anendothelial cell, or from infiltrating bone marrow include cells ofsolid tumors and liquid tumors (including hematological malignancies).Examples of solid tumors are described herein and include colorectalcancer, liver cancer, gastric cancer, lung cancer, brain cancer, kidneycancer, bladder cancer, thyroid cancer, prostate cancer, ovarian cancer,cervical cancer, uterine cancer, endometrial cancer, melanoma, breastcancer, and pancreatic cancer. Liquid tumors occur in the blood, bonemarrow, and lymph nodes and include leukemia (e.g., AML, ALL, CLL, andCML), lymphoma (e.g., Hodgkin lymphoma and non-Hodgkin lymphoma), andmyeloma (e.g., multiple myeloma). As used herein, the term cancer cellsinclude mature, progenitor, and cancer stem cells.

Bones are a common location for cancer to infiltrate once leaving theprimary tumor location. Once cancer resides in bone, it is frequently acause of pain to the individual. In addition, if the particular boneaffected is a source for production of blood cells in the bone marrow,the individual may develop a variety of blood cell related disorders.Breast and prostate cancer are examples of solid tumors that migrate tobones. Acute myelogenous leukemia (AML) and multiple myeloma (MM) areexamples of liquid tumors that migrate to bones. Cancer cells thatmigrate to bone will typically migrate to the endosteal region of thebone marrow. Once cancer cells have infiltrated into the marrow, thecells become quiescent and are protected from chemotherapy. Thecompounds of the present disclosure may block infiltration ofdisseminated cancer cells into bone marrow. A variety of subjects maybenefit from treatment with the compounds. Examples of such subjectsinclude individuals with a cancer type having a propensity to migrate tobone where the tumor is still localized or the tumor is disseminated butnot yet infiltrated bone, or where individuals with such a cancer typeare in remission.

The cancer patient population most likely to respond to treatment usingantagonists of E-selectin (e.g., compounds of Formula (I)) describedherein can be identified based on the mechanisms of action ofE-selectin. For example, patients may be selected that express a highlyactive E-selectin as determined by the genetic polymorphism forE-selectin of S128R (Alessandro et al., Int. J. Cancer 121:528-535,2007). In addition, patients for treatment by the compounds describedherein may also selected based on elevated expression of the E-selectinbinding ligands (sialyl Le^(a) and sialyl Le^(x)) as determined byantibodies directed against cancer-associated antigens CA-19-9 (Zheng etal., World J. Gastroenterol. 7:431-434, 2001) and CD65. In addition,antibodies HECA-452 and FH-6 which recognize similar carbohydrateligands of E-selectin may also be used in a diagnostic assay to selectthe cancer patient population most likely to respond to this treatment.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful in methods formobilizing cells from the bone marrow to the peripheral vasculature andtissues. As discussed herein, in some embodiments, the compounds andcompositions are useful for mobilizing hematopoietic cells, includinghematopoietic stem cells and hematopoietic progenitor cells. In someembodiments, the compounds act as mobilizing agents of normal blood celltypes. In some embodiments, the agents are used in methods formobilizing mature white blood cells (which may also be called leukocytesherein), such as granulocytes (e.g., neutrophils, eosinophils,basophils), lymphocytes, and monocytes from the bone marrow or otherimmune cell compartments such as the spleen and liver. Methods are alsoprovided for using the compounds of the present disclosure andpharmaceutical compositions comprising at least one such compound inmethods for mobilizing tumor cells from the bone marrow. The tumor cellsmay be malignant cells (e.g., tumor cells that are metastatic cancercells, or highly invasive tumor cells) in cancers. These tumor cells maybe of hematopoietic origin or may be malignant cells of another originresiding in the bone.

In some embodiments, the methods using the compounds described hereinare useful for mobilizing hematopoietic cells, such as hematopoieticstem cells and progenitor cells and leukocytes (including granulocytessuch as neutrophils), which are collected (i.e., harvested, obtained)from the subject receiving a compound of Formula (I) and at a later timeare administered back into the same subject (autologous donor) oradministered to a different subject (allogeneic donor). Hematopoieticstem cell replacement and hematopoietic stem cell transplantation havebeen successfully used for treating a number of diseases (includingcancers) as described herein and in the art. By way of example, stemcell replacement therapy or transplantation follows myeloablation of asubject, such as occurs with administration of high dose chemotherapyand/or radiotherapy. Desirably, an allogeneic donor shares sufficientHLA antigens with the recipient/subject to minimize the risk of hostversus graft disease in the recipient (i.e., the subject receiving thehematopoietic stem cell transplant). Obtaining the hematopoietic cellsfrom the donor subject (autologous or allogeneic) is performed byapheresis or leukapheresis. HLA typing of a potential donor and therecipient and apheresis or leukapheresis are methods routinely practicedin the clinical art.

By way of non-limiting example, autologous or allogenic hematopoieticstem cells and progenitors cells may be used for treating a recipientsubject who has certain cancers, such as Hodgkin lymphoma, non-Hodgkinlymphoma, or multiple myeloma. Allogeneic hematopoietic stem cells andprogenitors cells may be used, for example, for treating a recipientsubject who has acute leukemia (e.g., AML, ALL); chronic lymphocyticleukemia (CLL); amegakaryocytosis/congenital thrombocytopenia; aplasticanemia/refractory anemia; familial erythrophagocyticlymphohistiocytosis; myelodysplastic syndrome/other myelodysplasticdisorders; osteopetrosis; paroxysmal nocturnal hemoglobinuria; andWiskott-Aldrich syndrome, for example. Exemplary uses for autologoushematopoietic stem cells and progenitors cells include treating arecipient subject who has amyloidosis; germ cell tumors (e.g.,testicular cancer); or a solid tumor. Allogeneic hematopoietic stem celltransplants have also been investigated for use in treating solid tumors(see, e.g., Ueno et al., Blood 102:3829-36 (2003)).

In some embodiments of the methods described herein, the subject is nota donor of peripheral hematopoietic cells but has a disease, disorder,or condition for which mobilization of hematopoietic cells in thesubject will provide clinical benefit. Stated another way, while thisclinical situation is similar to autologous hematopoietic cellreplacement, the mobilized hematopoietic cells are not removed and givenback to the same subject at a later time as occurs, for example, with asubject who receives myeloablation therapy. Accordingly, methods areprovided for mobilizing hematopoietic cells, such as hematopoietic stemcells and progenitor cells and leukocytes (including granulocytes, suchas neutrophils), by administering at least once compound of Formula (I).Mobilizing hematopoietic stem cells and progenitor cells may be usefulfor treating an inflammatory condition or for tissue repair or woundhealing. See, e.g., Mimeault et al., Clin. Pharmacol. Therapeutics82:252-64 (2007).

In some embodiments, the methods described herein are useful formobilizing hematopoietic leukocytes (white blood cells) in a subject,which methods may be used in treating diseases, disorders, andconditions for which an increase in white blood cells, such asneutrophils, eosinophils, lymphocytes, monocytes, basophils, willprovide clinical benefit. For example, for cancer patients, thecompounds of Formula (I) are beneficial for stimulating neutrophilproduction to compensate for hematopoietic deficits resulting fromchemotherapy or radiation therapy. Other diseases, disorders, andconditions to be treated include infectious diseases and relatedconditions, such as sepsis. When the subject to whom at least onecompound of Formula (I) is administered is a donor, neutrophils may becollected for administration to a recipient subject who has reducedhematopoietic function, reduced immune function, reduced neutrophilcount, reduced neutrophil mobilization, severe chronic neutropenia,leucopenia, thrombocytopenia, anemia, and acquired immune deficiencysyndrome. Mobilization of mature white blood cells may be useful insubjects to improve or to enhance tissue repair, and to minimize orprevent vascular injury and tissue damage, for example following livertransplantation, myocardial infarction or limb ischemia. See, e.g.,Pelus, Curr. Opin. Hematol. 15:285-92 (2008); Lemoli et al.,Haematologica 93:321-24 (2008).

The compounds of Formula (I) may be used in combination with one or moreother agents that mobilize hematopoietic cells. Such agents include, forexample, G-CSF; AMD3100 or other CXCR4 antagonists; GRO-β (CXCL2) and anN-terminal 4-amino truncated form (SB-251353); IL-8SDF-1α peptideanalogs, CTCE-0021 and CTCE-0214; and the SDF1 analog, Met-SDF-1β (see,e.g., Pelus, supra and references cited therein). In some embodiments, acompound of Formula (I) may be administered with other mobilizing agentsused in the art, which may permit administration of a lower dose of GCSFor AMD3100, for example, than required in the absence of a compound ofFormula (I). The appropriate therapeutic regimen for administering acompound of Formula (I) in combination with another mobilizing agent oragents can be readily determined by a person skilled in the clinicalart.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful in methods forpreventing and/or treating mucositis. In some embodiments, at least onecompound of Formula (I) and/or a pharmaceutical composition comprisingat least one compound of Formula (I) may be used in methods describedherein for decreasing the likelihood of occurrence of mucositis in asubject who is in need thereof by administering the compound orcomposition to the subject. In some embodiments, the mucositis is chosenfrom oral mucositis, esophageal mucositis, and gastrointestinalmucositis. In some embodiments, the mucositis is alimentary mucositis.

It is believed that approximately half of all cancer patients undergoingtherapy suffer some degree of mucositis. Mucositis is believed to occur,for example, in virtually all patients treated with radiation therapyfor head and neck tumors, all patients receiving radiation along the GItract, and approximately 40% of those subjected to radiation therapyand/or chemotherapy for tumors in other locations (e.g., leukemias orlymphomas). It is also is believed to be highly prevalent in patientstreated with high dose chemotherapy and/or irradiation for the purposeof myeloablation, such as in preparation for stem cell or bone marrowtransplantation. The compounds of the present disclosure andpharmaceutical compositions comprising at least one such compound may beuseful in methods for treating and/or preventing mucositis in a subjectafflicted with cancer. In some embodiments, the subject is afflictedwith a cancer chosen from head and neck cancer, breast cancer, lungcancer, ovarian cancer, prostate cancer, lymphatic cancer, leukemiccancer, and/or gastrointestinal cancer. In some embodiments, themucositis is associated with radiation therapy and/or chemotherapy. Insome embodiments, the chemotherapy comprises administering atherapeutically effective amount of at least one compound chosen fromplatinum, cisplatin, carboplatin, oxaliplatin, mechlorethamine,cyclophosphamide, chlorambucil, azathioprine, mercaptopurine,vincristine, vinblastine, vinorelbine, vindesine, etoposide, teniposide,paclitaxel, docetaxel, irinotecan, topotecan, amsacrine, etoposide,etoposide phosphate, teniposide, 5-fluorouracil (5-FU), leucovorin,methotrexate, gemcitabine, taxane, leucovorin, mitomycin C,tegafur-uracil, idarubicin, fludarabine, mitoxantrone, ifosfamide anddoxorubicin.

In some embodiments, the method further comprises administering atherapeutically effective amount of at least one MMP inhibitor,inflammatory cytokine inhibitor, mast cell inhibitor, NSAID, NOinhibitor, or antimicrobial compound.

In some embodiments, the method further comprises administering atherapeutically effective amount of velafermin and/or palifermin.

In some embodiments, the method further comprises administering atherapeutically effective amount of Davanat®, mannose, and/or galactose.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful in methods fortreating and/or preventing thrombosis. As described herein methods areprovided for inhibiting formation of a thrombus or inhibiting the rateat which a thrombus is formed. These methods may therefore be used forpreventing thrombosis (i.e., reducing or decreasing the likelihood ofoccurrence of a thrombus in a statistically or clinically significantmanner).

Thrombus formation may occur in infants, children, teenagers and adults.An individual may have a hereditary predisposition to thrombosis.Thrombosis may be initiated, for example, due to a medical condition(such as cancer or pregnancy), a medical procedure (such as surgery) oran environmental condition (such as prolonged immobility). Otherindividuals at risk for thrombus formation include those who havepreviously presented with a thrombus.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful in methods fortreating individuals undergoing thrombosis or who are at risk of athrombotic event occurring. Such individuals may or may not have a riskof bleeding. In some embodiments, the individual has a risk of bleeding.In some embodiments, the thrombosis is a venous thromboembolism (VTE).VTE causes deep vein thrombosis and pulmonary embolism. Low molecularweight (LMW) heparin is the current mainstay therapy for the preventionand treatment of VTE. In many circumstances, however, the use of LMWheparin is contraindicated. LMW heparin is a known anti-coagulant anddelays clotting over four times longer than control bleeding times.Patients undergoing surgery, patients with thrombocytopenia, patientswith a history of stroke, and many cancer patients should avoidadministration of heparin due to the risk of bleeding. By contrast,administration of the compounds of Formula (I) significantly reduces thetime to clotting than occurs when LMW heparin is administered, and thusprovide a significant improvement in reducing bleeding time comparedwith LMW heparin. Accordingly, the compounds and pharmaceuticalcompositions described herein may not only be useful for treating apatient for whom the risk of bleeding is not significant, but also maybe useful in when the risk of bleeding is significant and the use ofanti-thrombosis agents with anti-coagulant properties (such as LMWheparin) is contraindicated.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be administered in combinationwith at least one additional anti-thrombosis agent. The compounds of thepresent disclosure and pharmaceutical compositions comprising at leastone such compound may function independently from the anti-thrombosisagent or may function in coordination with the at least oneanti-thrombosis agent. In addition, the administration of one or more ofthe compounds or compositions may be in conjunction with one or moreother therapies, e.g., for reducing toxicities of therapy. For example,at least one palliative agent to counteract (at least in part) a sideeffect of therapy may be administered. Agents (chemical or biological)that promote recovery and/or counteract side effects of administrationof antibiotics or corticosteroids are examples of such palliativeagents. The compounds of the present disclosure and pharmaceuticalcomposition comprising at least one such compound may be administeredbefore, after, or concurrently with administration of at least oneadditional anti-thrombosis agent or at least one palliative agent toreduce a side effect of therapy. Where administration is concurrent, thecombination may be administered from a single container or two (or more)separate containers.

The compounds of the present disclosure and pharmaceutical compositionscomprising at least one such compound may be useful for treating and/orpreventing at least one cardiovascular disease, disorder and/orcondition. Non-limiting examples of cardiovascular disease includeatherosclerosis, myocardial infarction, myocardial ischemia, coronaryartery stenosis (occlusion of the coronary arteries), chroniccardiovascular and/or arterial inflammation, acute cardiovascular and/orarterial inflammation, hypercholesterolemia, restenosis (narrowing ofthe vessel lumen), arrhythmia, thrombosis, hyperlipidemia, hypertension,dyslipoproteinemia, angina (cardiac chest pain), and vascularcomplications due to a cardiovascular disease (e.g., myocardialinfarction or myocardial ischemia).

In some embodiments, at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) may be administered prior to or subsequent to an acutecardiovascular event in the subject. In some embodiments, at least onecompound of Formula (I) and/or a pharmaceutical composition comprisingat least one compound of Formula (I) may be administered prior to orsubsequent to the development or diagnosis of a cardiovascular disease,disorder and/or condition in the subject. In some embodiments, the acutecardiovascular event is a myocardial infarction.

In some embodiments, a method for treatment and/or prevention ofatherosclerosis is disclosed, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I). Atherosclerosis generally describes a diseaseof the arterial blood vessels. As used herein, “atherosclerosis”includes, but is not limited to, chronic and/or acute atheroscleroticinflammation prior to or subsequent to the formation of at least oneatherosclerotic plaque in the subject. Atherosclerosis also includes,but is not limited to, chronic progressive atherosclerosis and/oratherosclerotic inflammation. Atherosclerosis also includes, but is notlimited to, acute atherosclerosis and/or atherosclerotic inflammationsubsequent to an acute vascular event in the subject (such as, forexample, myocardial infarction).

In some embodiments, at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) may be administered prior to or subsequent to the formation of atleast one atherosclerotic plaque, lesion or atheroma in the subject.

In some embodiments, the formation, progression, destabilization and/orrupture of at least one atherosclerotic plaque within the subject isreduced.

Atherosclerotic plaques may be characterized as stable or unstable(i.e., vulnerable to destabilization). Unstable atherosclerotic plaquesmay be susceptible to disruption or rupture, which exposes thrombogenicmaterial (i.e., thrombi) (e.g., collagen) to the circulation. This canproduce interruptions in blood flood (ischemia) in local or distalarteries, which can result in cardiovascular complications, such as, forexample, myocardial infarction (MI).

Destabilization of atherosclerotic plaques may occur via manymechanisms. Non-limiting examples of such mechanisms include superficialerosion of the endothelial cells that form the monolayer covering theintima, disruption of the microvessels that form in the atheroscleroticplaque, rupture (i.e., fracture) of the atherosclerotic plaque's fibrouscap, thinning or weakening of the fibrous cap (thus making itsusceptible to rupture), and the presence or increase in inflammatoryfactors that mediate destabilization. (Libby P., Nature, 420: 868-874(2002)).

A non-limiting example of inflammatory factors that mediatedestabilization is the presence of inflammatory cells. The progressionof atherosclerosis may be associated with systemically increasedinflammatory myeloid cells that are recruited to atheroscleroticplaques. (Murphy, A. J. et al., J. Clin. Invest., 121: 4138-4149 (2011);Averill, L. E. et al., Am. J. Pathol., 135: 369-377 (1989); Feldman, D.L. et al., Arterioscler. Thromb., 11: 985-994 (1991); Swirski, F. K. etal., J. Clin. Invest. 117: 195-205 (2007)). The presence of inflammatorymyeloid cells may be detrimental to a stable plaque. (Llodra, J. et al.,Proc. Natl. Acad. Sci. U.S.A., 101: 11779-11784 (2004)).

In some embodiments, the stability of at least one atheroscleroticplaque within the subject is increased. Non-limiting examples of stablefeatures of atherosclerotic plaques (i.e., stable phenotype) includesmaller plaque size, reduced (i.e., decreased, diminished, smaller)necrotic core size (measured by, for example, necrotic core area), and athicker fibrous cap of the atherosclerotic plaque. (See, e.g., Moore K.J. et al., Cell, 145: 341-355 (2011)).

In some embodiments, the size of at least one atherosclerotic plaquewithin the subject is decreased. In some embodiments, the necrotic coresize of at least one atherosclerotic plaque within the subject isdecreased. In some embodiments, the fibrous cap thickness of at leastone atherosclerotic plaque within the subject is increased.

In some embodiments, the administration of an effective amount of atleast one compound of Formula (I) and/or a pharmaceutical compositioncomprising at least one compound of Formula (I) reduces the levels ofextramedullary proliferation of hematopoietic stem and/or progenitorcells within the subject. In some embodiments, extramedullaryproliferation of hematopoietic stem and/or progenitor cells is reducedin the spleen and/or the liver. Non-limiting examples of extramedullaryproliferation of hematopoietic stem and/or progenitor cells includeextramedullary hematopoiesis and extramedullary myelopoiesis.

In some embodiments, the administration of an effective amount of atleast one compound of Formula (I) and/or a pharmaceutical compositioncomprising at least one compound of Formula (I) reduces the recruitmentand/or infiltration of myeloid cells to at least one atheroscleroticplaque within the subject. Non-limiting examples of myeloid cellsinclude monocytes, macrophages, neutrophils, basophils, eosinophils,erythrocytes, dendritic cells, and megakaryocytes and platelets.

In some embodiments, the at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) is administered after angioplasty, stenting procedure, atherectomy,bypass surgery, or other vessel-corrective techniques.

In some embodiments, the at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) is administered before angioplasty, stenting procedure, atherectomy,bypass surgery, or other vessel-corrective techniques.

In some embodiments, a method for treatment and prevention of myocardialinfarction is disclosed, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofFormula (I) and/or a pharmaceutical composition comprising at least onecompound of Formula (I). In some embodiments, the subject has previouslysuffered a myocardial infarction. In some embodiments, a compound ofFormula (I) may be administered before the occurrence of a myocardialinfarction in the subject. In some embodiments, a compound of Formula(I) may be administered after the occurrence of a first or subsequentmyocardial infarction in the subject.

In some embodiments, at least one compound of Formula (I) and/or apharmaceutical composition comprising at least one compound of Formula(I) is administered to the subject: within one (1) day of the subjectsuffering a myocardial infarction, within one (1) week of the subjectsuffering a myocardial infarction, within two (2) weeks of the subjectsuffering a myocardial infarction, within three (3) weeks of the subjectsuffering a myocardial infarction, within four (4) weeks of the subjectsuffering a myocardial infarction, within eight (8) weeks of the subjectsuffering a myocardial infarction, or within twelve (12) weeks of thesubject suffering a myocardial infarction.

The terms, “treat” and “treatment,” include medical management of adisease, disorder, and/or condition of a subject as would be understoodby a person of ordinary skill in the art (see, e.g., Stedman's MedicalDictionary). In general, an appropriate dose and treatment regimenprovide at least one of the compounds of the present disclosure in anamount sufficient to provide therapeutic and/or prophylactic benefit.For both therapeutic treatment and prophylactic or preventativemeasures, therapeutic and/or prophylactic benefit includes, for example,an improved clinical outcome, wherein the object is to prevent or slowor lessen an undesired physiological change or disorder, or to preventor slow or lessen the expansion or severity of such disorder. Asdiscussed herein, beneficial or desired clinical results from treating asubject include, but are not limited to, abatement, lessening, oralleviation of symptoms that result from or are associated with thedisease, condition, and/or disorder to be treated; decreased occurrenceof symptoms; improved quality of life; longer disease-free status (i.e.,decreasing the likelihood or the propensity that a subject will presentsymptoms on the basis of which a diagnosis of a disease is made);diminishment of extent of disease; stabilized (i.e., not worsening)state of disease; delay or slowing of disease progression; ameliorationor palliation of the disease state; and remission (whether partial ortotal), whether detectable or undetectable; and/or overall survival.“Treatment” can include prolonging survival when compared to expectedsurvival if a subject were not receiving treatment.

In some embodiments of the methods described herein, the subject is ahuman. In some embodiments of the methods described herein, the subjectis a non-human animal. Non-human animals that may be treated includemammals, for example, non-human primates (e.g., monkey, chimpanzee,gorilla, and the like), rodents (e.g., rats, mice, gerbils, hamsters,ferrets, rabbits), lagomorphs, swine (e.g., pig, miniature pig), equine,canine, feline, bovine, and other domestic, farm, and zoo animals.

The effectiveness of the compounds of the present disclosure in treatingand/or preventing diseases, disorders, and/or conditions treatable byinhibiting an activity of E-selectin can readily be determined by aperson of ordinary skill in the relevant art. Determining and adjustingan appropriate dosing regimen (e.g., adjusting the amount of compoundper dose and/or number of doses and frequency of dosing) can alsoreadily be performed by a person of ordinary skill in the relevant art.One or any combination of diagnostic methods, including physicalexamination, assessment and monitoring of clinical symptoms, andperformance of analytical tests and methods described herein, may beused for monitoring the health status of the subject.

Also provided herein are pharmaceutical compositions comprising at leastone compound of Formula (I). In some embodiments, the pharmaceuticalcompositions further comprise at least one additional pharmaceuticallyacceptable ingredient.

In pharmaceutical compositions, any one or more of the compounds of thepresent disclosure may be administered in the form of a pharmaceuticallyacceptable derivative, such as a salt, and/or it or they may also beused alone and/or in appropriate association, as well as in combination,with other pharmaceutically active compounds.

An effective amount or therapeutically effective amount refers to anamount of at least one compound of the present disclosure or apharmaceutical composition comprising at least one such compound that,when administered to a subject, either as a single dose or as part of aseries of doses, is effective to produce at least one therapeuticeffect. Optimal doses may generally be determined using experimentalmodels and/or clinical trials. Design and execution of pre-clinical andclinical studies for each of the therapeutics (including whenadministered for prophylactic benefit) described herein are well withinthe skill of a person of ordinary skill in the relevant art. The optimaldose of a therapeutic may depend upon the body mass, weight, and/orblood volume of the subject. In general, the amount of at least onecompound of Formula (I) as described herein, that is present in a dose,may range from about 0.1 mg to about 100 mg per kg weight of thesubject. The minimum dose that is sufficient to provide effectivetherapy may be used in some embodiments. Subjects may generally bemonitored for therapeutic effectiveness using assays suitable for thedisease, disorder and/or condition being treated or prevented, whichassays will be familiar to those having ordinary skill in the art andare described herein. The level of a compound that is administered to asubject may be monitored by determining the level of the compound (or ametabolite of the compound) in a biological fluid, for example, in theblood, blood fraction (e.g., serum), and/or in the urine, and/or otherbiological sample from the subject. Any method practiced in the art todetect the compound, or metabolite thereof, may be used to measure thelevel of the compound during the course of a therapeutic regimen.

The dose of a compound described herein may depend upon the subject'scondition, that is, stage of the disease, severity of symptoms caused bythe disease, general health status, as well as age, gender, and weight,and other factors apparent to a person of ordinary skill in the medicalart. Similarly, the dose of the therapeutic for treating a disease,disorder, and/or condition may be determined according to parametersunderstood by a person of ordinary skill in the medical art.

Pharmaceutical compositions may be administered in any mannerappropriate to the disease, disorder, and/or condition to be treated asdetermined by persons of ordinary skill in the medical arts. Anappropriate dose and a suitable duration and frequency of administrationwill be determined by such factors as discussed herein, including thecondition of the patient, the type and severity of the patient'sdisease, the particular form of the active ingredient, and the method ofadministration. In general, an appropriate dose (or effective dose) andtreatment regimen provides the composition(s) as described herein in anamount sufficient to provide therapeutic and/or prophylactic benefit(for example, an improved clinical outcome, such as more frequentcomplete or partial remissions, or longer disease-free and/or overallsurvival, or a lessening of symptom severity or other benefit asdescribed in detail above).

The pharmaceutical compositions described herein may be administered toa subject in need thereof by any one of several routes that effectivelydelivers an effective amount of the compound. Non-limiting examples ofsuitable administrative routes include topical, oral, nasal,intrathecal, enteral, buccal, sublingual, transdermal, rectal, vaginal,intraocular, subconjunctival, sublingual, and parenteral administration,including subcutaneous, intravenous, intramuscular, intrasternal,intracavernous, intrameatal, and intraurethral injection and/orinfusion.

The pharmaceutical compositions described herein may, for example, besterile aqueous or sterile non-aqueous solutions, suspensions, oremulsions, and may additionally comprise at least one pharmaceuticallyacceptable excipient (i.e., a non-toxic material that does not interferewith the activity of the active ingredient). Such compositions may, forexample, be in the form of a solid, liquid, or gas (aerosol).Alternatively, the compositions described herein may, for example, beformulated as a lyophilizate, or compounds described herein may beencapsulated within liposomes using technology known in the art. Thepharmaceutical compositions may further comprise at least one additionalpharmaceutically acceptable ingredient, which may be biologically activeor inactive. Non-limiting examples of such ingredients include buffers(e.g., neutral buffered saline or phosphate buffered saline),carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol,proteins, polypeptides, amino acids (e.g., glycine), antioxidants,chelating agents (e.g., EDTA and glutathione), stabilizers, dyes,flavoring agents, suspending agents, and preservatives.

Any suitable excipient or carrier known to those of ordinary skill inthe art for use in compositions may be employed in the compositionsdescribed herein. Excipients for therapeutic use are well known, and aredescribed, for example, in Remington: The Science and Practice ofPharmacy (Gennaro, 21^(st) Ed. Mack Pub. Co., Easton, Pa. (2005)). Ingeneral, the type of excipient may be selected based on the mode ofadministration, as well as the chemical composition of the activeingredient(s). Compositions may be formulated for the particular mode ofadministration. For parenteral administration, pharmaceuticalcompositions may further comprise water, saline, alcohols, fats, waxes,and buffers. For oral administration, pharmaceutical compositions mayfurther comprise at least one component chosen, for example, from any ofthe aforementioned ingredients, excipients and carriers, such asmannitol, lactose, starch, magnesium stearate, sodium saccharine,talcum, cellulose, kaolin, glycerin, starch dextrins, sodium alginate,carboxymethylcellulose, ethyl cellulose, glucose, sucrose, and magnesiumcarbonate.

The pharmaceutical compositions (e.g., for oral administration ordelivery by injection) may be in the form of a liquid. A liquidcomposition may include, for example, at least one the following: asterile diluent such as water for injection, saline solution, includingfor example physiological saline, Ringer's solution, isotonic sodiumchloride, fixed oils that may serve as the solvent or suspending medium,polyethylene glycols, glycerin, propylene glycol or other solvents;antibacterial agents; antioxidants; chelating agents; buffers and agentsfor the adjustment of tonicity such as sodium chloride or dextrose. Aparenteral preparation can be enclosed in ampoules, disposable syringesor multiple dose vials made of glass or plastic. In some embodiments,the pharmaceutical composition comprises physiological saline. In someembodiments, the pharmaceutical composition is an injectablecomposition, and in some embodiments, the injectable composition issterile.

For oral formulations, at least one of the compounds of the presentdisclosure can be used alone or in combination with at least oneadditive appropriate to make tablets, powders, granules and/or capsules,for example, those chosen from conventional additives, disintegrators,lubricants, diluents, buffering agents, moistening agents,preservatives, coloring agents, and flavoring agents. The pharmaceuticalcompositions may be formulated to include at least one buffering agent,which may provide for protection of the active ingredient from low pH ofthe gastric environment and/or an enteric coating. A pharmaceuticalcomposition may be formulated for oral delivery with at least oneflavoring agent, e.g., in a liquid, solid or semi-solid formulationand/or with an enteric coating.

Oral formulations may be provided as gelatin capsules, which may containthe active compound or biological along with powdered carriers. Similarcarriers and diluents may be used to make compressed tablets. Tabletsand capsules can be manufactured as sustained release products toprovide for continuous release of active ingredients over a period oftime. Compressed tablets can be sugar coated or film coated to mask anyunpleasant taste and protect the tablet from the atmosphere, or entericcoated for selective disintegration in the gastrointestinal tract.

A pharmaceutical composition may be formulated for sustained or slowrelease. Such compositions may generally be prepared using well knowntechnology and administered by, for example, oral, rectal orsubcutaneous implantation, or by implantation at the desired targetsite. Sustained-release formulations may contain the active therapeuticdispersed in a carrier matrix and/or contained within a reservoirsurrounded by a rate controlling membrane. Excipients for use withinsuch formulations are biocompatible, and may also be biodegradable; theformulation may provide a relatively constant level of active componentrelease. The amount of active therapeutic contained within a sustainedrelease formulation depends upon the site of implantation, the rate andexpected duration of release, and the nature of the condition to betreated or prevented.

The pharmaceutical compositions described herein can be formulated assuppositories by mixing with a variety of bases such as emulsifyingbases or water-soluble bases. The pharmaceutical compositions may beprepared as aerosol formulations to be administered via inhalation. Thepharmaceutical compositions may be formulated into pressurizedacceptable propellants such as dichlorodifluoromethane, propane,nitrogen and the like.

The compounds of the present disclosure and pharmaceutical compositionscomprising these compounds may be administered topically (e.g., bytransdermal administration). Topical formulations may be in the form ofa transdermal patch, ointment, paste, lotion, cream, gel, and the like.Topical formulations may include one or more of a penetrating agent orenhancer (also call permeation enhancer), thickener, diluent,emulsifier, dispersing aid, or binder. Physical penetration enhancersinclude, for example, electrophoretic techniques such as iontophoresis,use of ultrasound (or “phonophoresis”), and the like. Chemicalpenetration enhancers are agents administered either prior to, with, orimmediately following administration of the therapeutic, which increasethe permeability of the skin, particularly the stratum corneum, toprovide for enhanced penetration of the drug through the skin.Additional chemical and physical penetration enhancers are described in,for example, Transdermal Delivery of Drugs, A. F. Kydonieus (ED) 1987CRL Press; Percutaneous Penetration Enhancers, eds. Smith et al. (CRCPress, 1995); Lenneräs et al., J. Pharm. Pharmacol. 54:499-508 (2002);Karande et al., Pharm. Res. 19:655-60 (2002); Vaddi et al., Int. J.Pharm. 91:1639-51 (2002); Ventura et al., J. Drug Target 9:379-93(2001); Shokri et al., Int. J. Pharm. 228(1-2):99-107 (2001); Suzuki etal., Biol. Pharm. Bull. 24:698-700 (2001); Alberti et al., J. ControlRelease 71:319-27 (2001); Goldstein et al., Urology 57:301-5 (2001);Kiijavainen et al., Eur. J. Pharm. Sci. 10:97-102 (2000); and Tenjarlaet al., Int. J. Pharm. 192:147-58 (1999).

Kits comprising unit doses of at least one compound of the presentdisclosure, for example in oral or injectable doses, are provided. Suchkits may include a container comprising the unit dose, an informationalpackage insert describing the use and attendant benefits of thetherapeutic in treating the pathological condition of interest, and/oroptionally an appliance or device for delivery of the at least onecompound of Formula (I) and/or pharmaceutical composition comprising thesame.

EXAMPLES

Unless otherwise noted, all chemicals were reagent grade materials fromcommercial suppliers and were used without further purification. NMRdata were recorded on a Bruker 300 MHz spectrometer or a Bruker 400 MHzspectrometer and the chemical shifts are reported in ppm. Couplingconstants (J) are reported in hertz. Mass spectra were recorded on anAgilent 1100 Series LC/MSD spectrometer.

Example 1 Synthesis of Compound 17

Compound 3: A solution of compound 3 was prepared according toBioconjugate. Chem. 2014, 25, 1444-52.

Compound 4: Sulfur trioxide pyridine complex was purified by placing thematerial in a fritted funnel and washing with ice water until the pH ofthe filtrate was between 5-6. The material was then dried by washingwith ice cold ethanol followed by dichloromethane and then ether. Thematerial was used immediately for reaction.

3-Azidopropanol (0.56 g, 5.6 mmol) was dissolved in dry pyridine (20 mL)under an argon atmosphere. Freshly purified sulfur trioxide pyridinecomplex (2.92 g, 18.3 mmol) was added and the reaction mixture wasstirred overnight at 60° C. The solvent was removed and the resultingsolid was dissolved in 10 mL de-ionized water. The solution was cooledon an ice bath and the pH was adjusted to ˜9 by slow addition of 3NNaOH. The solvent was removed and the resulting solid was suspended inisopropanol and filtered to afford 410 mg compound 4 as a yellow-brownsolid. MS negative mode: m/z=180.0 [M−Na]⁻ C₃H₆N₃NaO₄S (203).

¹H NMR (400 MHz, D₂O) δ 4.04 (t, J=6.0 Hz, 2H), 3.37 (t, J=6.6 Hz, 2H),1.86 (p, J=6.3 Hz, 2H).

Compound 6: Prepared according to Bioorg. Med. Chem. Lett. 1995, 5,2321-2324 starting with D-threonolactone.

Compound 7: Compound 6 (500 mg, 1 mmol) was dissolved in 9 mLacetonitrile. Potassium hydroxide (1 mL of a 2M solution) was added andthe reaction mixture was stirred at 50° C. for 12 hours. The reactionmixture was partitioned between dichloromethane and water. The phaseswere separated and the aqueous phase was extracted 3 times withdichloromethane. The aqueous phase was acidified with 1N HCl until pH ˜1and extracted 3 times with dichloromethane. The combined dichloromethaneextracts from after acidification of the aqueous phase were concentratedin vacuo to give compound 7 as a yellow oil (406 mg). LCMS (C-18; 5-95H₂O/MeCN): UV (peak at 4.973 min), positive mode: m/z=407 [M+H]⁺;negative mode: m/z=405 [M−H]⁻C₂₅H₂₆O₅ (406).

Compound 11: Compound 47 (prepared according to WO 2013/096926) (0.18 g)was dissolved in ethylenediamine (2 mL) and stirred at 80° C. for 8hours. The solvent was removed in vacuo and the crude material separatedby C-18 reverse phase chromatography to afford 0.15 g of compound 11. MSpositive mode: m/z=756.3 [M+H]⁺. MS negative mode: m/z=732.3 [M−Na]⁻C₃₄H₅₈N₃NaO₁₄ (755.38).

¹H NMR (400 MHz, D₂O) δ 5.02 (d, J=3.9 Hz, 1H), 4.89 (q, J=6.6 Hz, 1H),4.54 (d, J=8.5 Hz, 1H), 3.95 (t, J=8.1 Hz, 1H), 3.89-3.85 (m, 3H),3.83-3.74 (m, 2H), 3.72 (d, J=5.8 Hz, 2H), 3.55 (t, J=5.9 Hz, 1H),3.50-3.44 (m, 3H), 3.33 (t, J=9.7 Hz, 1H), 3.11 (t, J=6.1 Hz, 2H), 2.36(t, J=12.6 Hz, 1H), 2.17 (br d, J=12.0 Hz, 1H), 2.04 (s, 3H), 1.90 (d,J=13.7 Hz, 1H), 1.87-1.83 (m, 1H), 1.76-1.66 (m, 2H), 1.64-1.41 (m, 3H),1.41-1.28 (m, 1H), 1.22 (d, J=6.4 Hz, 3H), 1.20-1.14 (m, 3H), 0.92-0.87(m, 2H), 0.84 (t, J=7.3 Hz, 3H).

Compound 12: To a stirred and cooled (15° C.) solution of propiolic acid(82 μl, 1.19 mmol) in DCM (5 mL) under argon was added oxalyl chloride(100 μl, 1.19 mmol) and DMF (5 μl, cat). The resulting mixture isstirred for 20 min before addition to a solution of compound 11 (200 mg,0.27 mmol) in MeOH (20 mL) containing saturated sodium bicarbonatesolution (4 mL). The resulting mixture was stirred at room temperaturefor 30 min; LCMS at that time showed the reaction was complete. Thereaction was filtered and partially concentrated under reduced pressureto the water fraction, which, was in-turn freeze-dried. Gel permeationof the residue (water as eluent) afforded compound 12 as a white solid(214 mg) (100%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.342 min),UV (peak at 3.247 min), negative mode: m/z=785.2 [M−H]⁻ C₃₇H₅₉N₃O₁₅(786).

¹H NMR (300 MHz, D₂O) δ 4.96 (d, J=3.9 Hz, 1H), 4.83 (q, J=8.0 Hz, 1H),4.45 (d, J=8.6 Hz, 1H), 3.93 (t, J=9.5 Hz, 1H), 3.84-3.78 (m, 3H),3.78-3.72 (m, 2H), 3.72-3.62 (m, 3H), 3.49 (t, J=5.9 Hz, 1H), 3.42-3.32(m, 2H), 3.32-3.21 (m, 4H), 2.25 (br t, J=12.1 Hz, 1H), 2.09 (br d,J=7.3 Hz, 1H), 2.00 (s, 3H), 1.80 (br d, J=10.3 Hz, 2H), 1.74-1.37 (m,7H), 1.37-1.20 (m, 3H), 1.17 (d, J=6.5 Hz, 3H), 1.14-1.01 (m, 3H),0.93-0.83 (m, 2H), 0.79 (t, J=7.3 Hz, 3H).

Compound 17: A stirred solution of compound 12 (75 mg, 95 μmol) andazide 3 (288 mg, 114 μmol) in HPLC grade water (6 ml) under argon wastreated with a pre-mixed solution of copper sulfate and THPTA (19 μmol,0.5 mL of 40 mmol aqueous solution) and sodium ascorbate (124 μmol,0.125 mL of 1M solution). After 2 h the reaction was lyophilized. Gelpermeation of the residue (water as eluent) afforded compound 17 as anoff-white solid (46 mg, 39%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at3.046 min), UV (peak at 2.846/2.952 min), negative mode: m/z=592[(M−4Na)/2]⁻ C₄₂H₆₆N₆Na₃O₂₇S₃ (1274.27).

¹H NMR (300 MHz, D₂O) δ 8.47 (s, 1H), 4.95 (d, J=4.0 Hz, 1H), 4.85 (q,J=7.0 Hz, 1H), 4.43 (d, J=8.5 Hz, 1H), 4.15 (br s, 1H), 4.01 (s, 6H),3.93 (br s, 1H), 3.81 (dd, J=10.5, 3.2 Hz, 1H), 3.74-3.69 (m, 2H),3.69-3.57 (m, 3H), 3.54-3.41 (m, 1H), 3.41-3.30 (m, 1H), 3.24 (t, J=9.6Hz, 1H), δ 2.22 (br t, J=12.4 Hz, 1H), 2.06 (br s, 1H), 1.99 (s, 3H),1.84-1.61 (m, 2H), 1.60-1.39 (m, 7H), 1.26 (q, J=12.7, 12.2 Hz, 1H),1.16 (d, J=6.5 Hz, 3H), 0.96 (dd, J=68.9, 12.0 Hz, 1H), 0.74 (t, J=7.0Hz, 3H).

Example 2 Synthesis of Compound 18

Compound 13: Compound 11 (501 mg, 0.66 mmol, prepared according to WO2013/096926) and triethylamine (0.18 mL, 1.32 mmol) were dissolved in 5mL MeOH and cooled on an ice bath. Ethyl trifluoroacetate (0.1 mL, 0.79mmol) was added dropwise and the reaction mixture was stirred 1 hour onthe ice bath. The solvent was removed and the residue was azeotropedthree times from toluene to afford compound 13. The crude material wasused without further purification. MS m/z=852.3 [M+H]: C₃₆H₅₇F₃N₃NaO₁₅.

¹H NMR (400 MHz, DMSO-d6) δ 9.41 (t, J=5.4 Hz, 1H), 7.94 (t, J=5.4 Hz,1H), 7.63 (br s, 1H), 4.76 (q, J=6.5 Hz, 1H), 4.67 (d, J=3.5 Hz, 1H),4.44 (br s, 2H), 4.16 (d, J=6.5 Hz, 2H), 4.11 (d, J=4.4 Hz, 1H), 3.68(br s, 1H), 3.64 (d, J=8.7 Hz, 1H), 3.58-3.47 (m, 3H), 3.45-3.37 (m,2H), 3.28-3.19 (m, 3H), 2.89 (br s, 4H), 2.11 (br t, I=12.05 Hz, 1H),2.04 (br d, J=12.3 Hz, 1H), 1.99-1.90 (m, 1H), 1.77 (s, 3H), 1.72-1.56(m, 2H), 1.52 (br d, J=13.1 Hz, 1H), 1.46-1.36 (m, 3H), 1.20 (q, J=12.1Hz, 2H), 1.06 (d, J=6.3 Hz, 3H), 1.04-0.84 (m, 2H), 0.79 (t, J=7.4 Hz,3H).

Compound 14: Crude compound 13 (0.66 mmol) was dissolved in dry DMF andcooled on an ice bath under an argon atmosphere. Diisopropylethylamine(0.23 mL, 1.32 mmol) was added followed by HATU (300 mg, 0.79 mmol). Thereaction mixture was stirred 15 minutes. Azetidine (90 μL, 1.32 mmol)was added and the reaction mixture was stirred overnight allowing towarm to room temperature during which time the product precipitated fromthe solution. The reaction mixture was suspended in acetonitrile,stirred 1 minute and the mixture allowed to settle. The yellowacetonitrile solution was carefully removed using a pipet. Thisprocedure was repeated until the acetonitrile solution was no longeryellow then 2 more times. The remaining solid was dried in vacuo toafford 502 mg compound 14 (87% yield for 2 steps). MS m/z=891.3 [M+Na]:C₃₉H₆₃F₃N₄NaO₁₄.

¹H NMR (400 MHz, DMSO-d6) δ 9.40 (t, J=2.5 Hz, 1H), 7.93 (t, J=6.1 Hz,1H), 7.70 (br s, 1H), 4.75 (q, J=6.7 Hz, 1H), 4.67 (d, J=3.3 Hz, 1H),4.55 (t, J=5.5 Hz, 1H), 4.31 (d, J=2.9 Hz, 1H), 4.26 (q, J=7.9 Hz, 1H),4.19-4.14 (m, 2H), 4.13-4.09 (m, 2H), 3.94 (br d, J=9.5 Hz, 1H), 3.87(q, J=7.5 Hz, 1H), 3.63 (s, 1H), 3.56-3.49 (m, 2H), 3.46-3.36 (m, 3H),3.29-3.18 (m, 3H), 3.14-3.03 (m, 2H), 2.22 (p, J=7.7 Hz, 2H), 2.11 (brt, J=12.1 Hz, 0H), 2.02 (br d, J=13.1 Hz, 1H), 1.98-1.90 (m, 1H), 1.79(s, 3H), 1.74-1.50 (m, 4H), 1.49-1.34 (m, 1H), 1.29-1.08 (m, 2H), 1.05(d, J=6.4 Hz, 3H), 1.02-0.84 (m, 2H), 0.79 (t, J=7.4 Hz, 3H).

Compound 15: Compound 14 (498 mg, 0.57 mmol) was suspended in 5 mL MeOHand cooled on an ice bath. Sodium methoxide (0.15 mL of a 25 wt % inMeOH solution, 0.68 mmol) was added and the reaction mixture was stirredallowing to warm to room temperature. The reaction mixture slowly becamehomogeneous. After stirring overnight at room temperature, MS indicatescompound 13 is still present. Sodium methoxide solution (50 μL) wasadded and the reaction mixture was stirred at room temperature foranother 5 hours. The reaction mixture was quenched by addition of 0.5 mLHOAc. The volatiles were removed in vacuo and the residue was separatedby reverse phase chromatography using a 10 g C-18 cartridge and elutingwith water then 3/1 water/MeOH then 1/1 water/MeOH. The productcontaining fractions were concentrated and the residue lyophilized toafford 420 mg of compound 15 as an off-white solid (94% yield). MSm/z=773.4 [M+H]: C₃₇H₆₄N₄O₁₃.

¹H NMR (400 MHz, DMSO-d6) δ 7.91 (t, I=5.4 Hz, 1H), 4.67 (q, J=6.5 Hz,1H), 4.59 (d, J=3.4 Hz, 1H), 4.25 (s, 1H), 4.20 (q, J=7.9 Hz, 1H), 4.04(q, J=8.0 Hz, 1H), 3.86 (dd, J=10.2, 2.6 Hz, 1H), 3.81 (q, J=8.4 Hz,1H), 3.56 (s, 1H), 3.51-3.41 (m, 3H), 3.40-3.28 (m, 5H), 3.19-3.11 (m,2H), 3.10 (s, 3H), 2.99 (dd, J=11.2, 7.8 Hz, 1H), 2.68 (t, J=6.5 Hz,2H), 2.43 (p, J=1.8 Hz, 3H), 2.15 (q, J=7.7 Hz, 1H), 2.11-2.01 (m, OH),2.01-1.83 (m, 1H), 1.71 (s, 3H), 1.68-1.50 (m, 2H), 1.50-1.26 (m, 2H),1.20-1.02 (m, 3H), 1.00-0.95 (m, 1H), 0.94-0.76 (m, OH), 0.72 (t, J=7.2Hz, 1H).

Compound 16: Propiolic acid (200 μl, 3.23 mmol) was dissolved in DCM (5mL) under argon and cooled on an ice bath. Oxalyl chloride (1.6 ml of a2M solution in DCM, 3.2 mmol) and DMF (1 drop, cat) were added and thereaction mixture was stirred for 1 hour. Compound 15 (210 mg, 0.27 mmol)was dissolved in 5 mL of saturated aqueous sodium bicarbonate solutionand cooled on an ice bath. The solution of propiolyl chloride was addedand the reaction mixture was stirred 3 hours on an ice bath. Volatileswere removed in vacuo and the resulting aqueous reaction mixture wasseparated by reverse phase chromatography using a 10 g C-18 cartridgeand eluting with water followed by 1/1 water/MeOH followed by ⅓water/MeOH to give recovered compound 15 (in the 1/1 water/MeOH eluent)and partially purified product (in the ⅓ water/MeOH eluent). Thepartially purified product was further purified using a 5 g C-18cartridge and eluting with water followed by 1/1 water/MeOH followed by⅓ water/MeOH to afford 29 mg compound 16 as a white solid (13%). MS,positive mode: m/z=847.4 [M+Na] C₄₀H₆₄N₄O₁₄ (824).

¹H NMR (400 MHz, D₂O) δ 4.93 (d, J=4.0 Hz, 1H), 4.79 (q, J=6.7 Hz, 1H),4.42 (d, J=8.6 Hz, 1H), 4.24 (q, J=8.7 Hz, 1H), 4.15 (q, J=8.7 Hz, 1H),4.01 (dd, J=9.9, 2.4 Hz, 1H), 3.94 (q, J=8.2 Hz, 1H), 3.87 (d, J=11.3Hz, 0H), 3.77 (dd, J=10.7, 3.3 Hz, 1H), 3.74-3.65 (m, 3H), 3.65-3.56 (m,2H), 3.49-3.37 (m, 1H), 3.36-3.28 (m, 1H), 3.28-3.16 (m, 3H), 2.29-2.16(m, 3H), 2.03 (d, J=8.0 Hz, 1H), 1.95 (s, 3H), 1.76 (d, J=13.3 Hz, 2H),1.61 (q, J=12.9 Hz, 1H), 1.54-1.38 (m, 2H), 1.33-1.16 (m, 2H), 1.12 (d,J=6.5 Hz, 3H), 1.10-1.01 (m, 1H), 0.85 (q, J=10.4, 9.8 Hz, 1H), 0.76 (t,J=7.3 Hz, 3H).

Compound 18: Using an analogous procedure to the procedure in FIG. 5,compound 18 was prepared from compound 16 and azide 3. MS negative mode:m/z=407.6 [(M−3Na)/3]⁻ C₄₅H₇₂N₇Na₃O₂₆S₃ (1291).

¹H NMR (400 MHz, D₂O) δ 8.36 (s, 1H), 4.86 (d, J=4.0 Hz, 1H), 4.77 (q,J=6.7 Hz, 1H), 4.59 (s, 1H), 4.35 (d, J=8.2 Hz, 1H), 4.28 (q, J=8.7 Hz,1H), 4.09 (q, J=8.7 Hz, 1H), 4.02-3.98 (m, 1H), 3.98-3.81 (m, 6H), 3.72(dd, J=10.5, 3.3 Hz, 1H), 3.68 (d, J=2.7 Hz, 1H), 3.66-3.58 (m, 2H),3.58-3.49 (m, 3H), 3.46-3.35 (m, 2H), 3.26-3.13 (m, 2H), 3.08 (br t,J=11.6 Hz, 1H), 2.28-2.10 (m, 1H), 2.05-1.94 (m, 1H), 1.93 (s, 3H),1.75-1.55 (m, 2H), 1.54-1.29 (m, 3H), 1.26-1.10 (m, 2H), 1.07 (d, J=6.5Hz, 3H), 1.05-0.70 (m, 2H), 0.67 (t, J=7.3 Hz, 3H).

Example 3 Synthesis of Compound 19

Compound 4: Sulfur trioxide pyridine complex was purified by placing thematerial in a fritted funnel and washing with ice water until the pH ofthe filtrate was between 5-6. The material was then dried by washingwith ice cold ethanol followed by dichloromethane and then ether. Thematerial was used immediately for reaction.

3-Azidopropanol (0.56 g, 5.6 mmol) was dissolved in dry pyridine (20 mL)under an argon atmosphere. Freshly purified sulfur trioxide pyridinecomplex (2.92 g, 18.3 mmol) was added and the reaction mixture wasstirred overnight at 60° C. The solvent was removed and the resultingsolid was dissolved in 10 mL de-ionized water. The solution was cooledon an ice bath and the pH was adjusted to ˜9 by slow addition of 3NNaOH. The solvent was removed and the resulting solid was suspended inisopropanol and filtered to afford 410 mg compound 4 as a yellow-brownsolid. MS negative mode: m/z=180.0 [M−Na]⁻ C₃H₆N₃NaO₄S (203).

¹H NMR (400 MHz, D₂O) δ 4.04 (t, J=6.0 Hz, 2H), 3.37 (t, J=6.6 Hz, 2H),1.86 (p, J=6.3 Hz, 2H).

Compound 19: Compound 19 was prepared in a procedure analogous to theprocedure in FIG. 5, using compound 16 and compound 4 as startingmaterials. MS negative mode: m/z=1004.4 [(M−Na)]⁻ C₄₃H₇₀N₇NaO₁₈S (1027).

¹H NMR (400 MHz, D₂O) δ 8.36 (s, 1H), 4.95 (d, J=4.0 Hz, 1H), 4.83 (q,J=8.0 Hz, 1H), 4.57 (t, J=8.0 Hz, 2H), 4.44 (broad d, J=8.0 Hz, 1H),4.31 (q, J=8.0 Hz, 1H), 4.19 (q, J=8.0 Hz, 1H), 4.11-4.04 (m, 1H),4.03-3.86 (m, 5H), 3.80 (dd, J=12.0, 4.0 Hz, 1H), 3.76 (broad d, 1H),3.72 (broad d, 2H), 3.69-3.54 (m, 4H), 3.53-3.45 (m, 2H), 3.43-3.32 (m,3H), 3.24 (br t, J=8.0 Hz, 1H), 2.36-2.17 (m, 5H), 2.12-2.03 (m, 1H),1.97 (s, 3H), 1.83-1.40 (m, 9H), 1.37-0.97 (m, J=8.0 Hz, 11H), 0.96-0.75(m, 3H), 0.71 (t, J=8.0 Hz, 3H).

Example 4 Synthesis of Compound 20

Compound 20: Using an analogous procedure to the procedure in FIG. 5,compound 20 was prepared from compound 16 and azide 2. MS positive mode:m/z=1008.3 [(M+Na)]⁻ C₄₅H₇₅N₇O₁₇ (985).

¹H NMR (400 MHz, D₂O) δ 8.30 (s, 1H), 4.89 (d, J=4.0 Hz, 1H), 4.77 (q,J=8.0 Hz, 1H), 4.46-4.32 (m, 3H), 4.24 (q, J=8.0 Hz, 1H), 4.15 (q, J=8.0Hz, 1H), 4.05-3.98 (m, 1H), 3.94 (q, J=8.0 Hz, 2H), 3.89-3.73 (m, 1H),3.75 (dd, J=12.0, 4.0 Hz, 1H), 3.72-3.70 (m, 1H), 3.69-3.64 (m, 2H),3.63-3.55 (m, 3H), 3.54-3.47 (m, 1H), 3.46-3.31 (m, 9H), 3.30-3.21 (m,1H), 3.17 (br t, J=12.0 Hz, 1H), 2.30-2.11 (m, 3H), 2.07-1.97 (m, 1H),1.91 (s, 3H), 1.78-1.31 (m, 9H), 1.30-1.19 (m, 3H), 1.18-1.00 (m, J=8.0Hz, 7H), 0.99-0.70 (m, 4H), 0.66 (t, J=8.0 Hz, 3H).

Example 5 Synthesis of Compound 21

Compound 21: Using an analogous procedure to the procedure in FIG. 5,compound 20 was prepared from compound 16 and azide 3-azidopropanol. MSpositive mode: m/z=948.3 [(M+Na)]⁻ C₄₃H₇₁N₇O₁₅ (925).

¹H NMR (400 MHz, D₂O) δ 8.31 (s, 1H), 7.82 (s, 1H), 4.89 (d, J=4.0 Hz,1H), 4.78 (q, J=8.0 Hz, 1H), 4.47 (t, J=8.0 Hz, 2H), 4.39 (t, J=8.0 Hz,2H), 4.24 (q, J=8.0 Hz, 1H), 4.15 (q, J=8.0 Hz, 1H), 4.05-3.99 (m, 1H),3.94 (q, J=8.0 Hz, 2H), 3.89-3.79 (m, 1H), 3.79-3.64 (m, 6H), 3.63-3.54(m, 3H), 3.53-3.33 (m, 8H), 3.31-3.22 (m, 1H), 3.17 (br t, J=12.0 Hz,1H).

Example 6 Prophetic Synthesis of Compound 55

Compound 51: Compound 51 can be prepared in an analogous fashion to FIG.3 by substituting 2-(proparglyoxy)ethanamine for ethylenediamine.

Compound 55: Compound 55 can be prepared by the same procedure seen inFIG. 5 by substituting compound 51 for compound 12.

Example 7 Prophetic Synthesis of Compound 56

Compound 56: Compound 56 can be prepared by the same procedure seen inFIG. 5 substituting compound 51 for compound 12 and substitutingpentyn-5-ol for compound 3.

Example 8 Prophetic Synthesis of Compound 57

Compound 53: Compound 53 can be prepared in an analogous fashion to FIG.4 by substituting 6-(2-propyn-1-yloxy)-3-pyridinecarboxylic acid forpropiolic acid.

Compound 57: Compound 57 can be prepared by the same procedure seen inFIG. 5 substituting compound 53 for compound 12 and substitutingcompound 4 for compound 3.

Example 9 Prophetic Synthesis of Compound 58

Compound 54: Compound 54 can be prepared in an analogous fashion to FIG.4 by substituting 5-ethynyl-3-pyridinecarboxylic acid for propiolicacid.

Compound 58: Compound 58 can be prepared by the same procedure as seenin FIG. 5 by substituting compound 54 for compound 12 and substituting3-azidopropanol for compound 3.

Example 10 Synthesis of Compound 24

Compound 6: Prepared according to Bioorg. Med. Chem. Lett. 1995, 5,2321-2324 starting with D-threonolactone.

Compound 7: Compound 6 (500 mg, 1 mmol) was dissolved in 9 mLacetonitrile. Potassium hydroxide (1 mL of a 2M solution) was added andthe reaction mixture was stirred at 50° C. for 12 hours. The reactionmixture was partitioned between dichloromethane and water. The phaseswere separated and the aqueous phase was extracted 3 times withdichloromethane. The aqueous phase was acidified with 1N HCl until pH ˜1and extracted 3 times with dichloromethane. The combined dichloromethaneextracts from after acidification of the aqueous phase were concentratedin vacuo to give compound 7 as a yellow oil (406 mg). LCMS (C-18; 5-95H₂O/MeCN): UV (peak at 4.973 min), positive mode: m/z=407 [M+H]⁺;negative mode: m/z=405 [M−H]⁻C₂₅H₂₆O₅ (406).

Compound 22: HATU (52 mg, 0.14 mmol) and DIPEA (28 μL, 0.16 mmol) wereadded to a solution of compound 7 (54 mg, 0.13 mmol) in DMF (5 mL)cooled to 0° C. The mixture was stirred at 0° C. for 5 min. A solutionof compound 11 (78 mg, 0.106 mmol) in water (2 mL) was then added andthe yellow solution was stirred at 0° C.→r.t. for 12 h. The volatileswere evaporated and co-evaporated with toluene (2×). The resultingyellow residue was dried under high vacuum for 3 h. To a solution of thelatter residue in pyridine (4 mL) was added acetic anhydride (2 mL) andthe solution was stirred at r.t. for 12 h. The volatiles were evaporatedand co-evaporated with toluene (2×). The resulting yellow material waspurified by column chromatography (DCM/MeOH 0→5%) to give compound 22 asa yellow solid (141 mg). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 5.526min); UV (peak at 5.432 min).

Compound 23: A suspension of compound 22 (101 mg, 0.08 mmol) and Pd/C10% (5 mg) in MeOH (10 mL) was degassed (3×) prior to the addition ofhydrogen using a balloon. The mixture was stirred at r.t. for 12 h. LCMSanalysis shows reaction not complete. The reaction was stirred forfurther 12 h. LCMS analysis shows reaction gone to completion. Thecatalyst was filtered off through a short pad of celite and the filtrateconcentrated under vacuum to give compound 23 as a white glass (83 mg).LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 4.164 min).

Compound 24: Sulfur trioxide pyridine complex was purified as describedin the procedure for compound 4. A solution of the purified sulfatingreagent (132 mg, 0.83 mmol) and compound 23 (83 mg, 0.08 mmol) inpyridine (3 mL) was stirred at 67° C. for 1 h. The reaction mixture wasconcentrated under vacuum. The resulting yellow solid was dissolved inwater and cooled to 0° C. A 1N solution of NaOH was then added slowlyuntil pH-10 and the latter was freeze dried. The resulting white solidwas then dissolved in MeOH, cooled to −15° C. and treated with NaOMe 25%solution in MeOH (0.5 mL). The mixture was stirred between −15° C. and−10° C. for 30 min. The reaction mixture was neutralised by adding HCl1N until pH-7. The volatiles were evaporated and the resulting yellowresidue was purified by Gel Permeation (water as eluent). The collectedfractions were lyophilised to give compound 24 as an off-white solid (50mg, 61% over 2 steps). MS negative mode: m/z=544.4 [(M−4Na)/2]⁻C₃₈H₆₁N₃Na₄O₂₇S₃ (1180).

¹H NMR (300 MHz, D₂O) δ 4.98 (d, J=4.0 Hz, 1H), 4.91-4.81 (m, 2H), 4.47(d, J=8.2 Hz, 1H), 4.28 (dd, J=10.7, 5.0 Hz, 1H), 4.18 (dd, J=10.7, 7.3Hz, 1H), 3.98 (t, J=9.4 Hz, 1H), 3.88 (s, 1H), 3.84 (dd, J=10.7, 3.1 Hz,1H), 3.77-3.73 (m, 1H), 3.72-3.69 (m, 1H), 3.67 (d, J=5.6 Hz, 2H, 3.54(t, J=5.8 Hz, 1H), 3.46 (d, J=8.6 Hz, 1H), 3.42-3.32 (m, 2H), 3.27 (t,J=9.6 Hz, 2H), 2.31 (t, J=12.4 Hz, 1H), 2.18 (d, J=11.6 Hz, 1H), 2.02(s, 3H), 1.87-1.73 (m, 3H), 1.72-1.62 (m, 2H), 1.62-1.40 (m, 5H),1.41-1.22 (m, 2H), 1.18 (d, J=6.5 Hz, 3H), 0.96-0.85 (m, 1H), 0.81 (t,J=7.2 Hz, 3H).

Example 11 Synthesis of Compound 25

Compound 8: Prepared in an analogous fashion to compound 7 usingL-erythronolactone as the starting material. LCMS (C-18; 5-95 H₂O/MeCN):ELSD (5.08 min), UV (peak at 4.958 min), positive mode: m/z=407 [M+H]⁺;negative mode: m/z=405 [M−H]⁻ C₂₅H₂₆O₅ (406).

Compound 25: Compound 25 was prepared using an analogous sequence tocompound 24, substituting compound 8 for compound 7. MS negative mode:m/z=544.8 [(M−4Na)/2]⁻ C₃₈H₆₁N₃Na₄O₂₇S₃ (1180).

¹H NMR (300 MHz, D₂O) δ 4.97 (d, J=4.0 Hz, 1H), 4.95 (d, J=4.0 Hz, 1H),4.88-4.78 (m, 2H), 4.47 (d, J=8.6 Hz, 1H), 4.26 (dd, J=10.7, 5.9 Hz,1H), 4.18 (dd, J=10.6, 5.2 Hz, 1H), 3.97 (dd, J=10.4, 8.5 Hz, 1H),3.87-3.77 (m, 3H), 3.77-3.71 (m, 2H), 3.71-3.61 (m, 3H), 3.53 (t, J=5.9Hz, 1H), 3.47-3.36 (m, 1H), 3.36-3.20 (m, 4H), 2.29 (t, J=12.4 Hz, 1H),2.15 (d, J=12.4 Hz, 1H), 2.01 (s, 3H), 1.88-1.47 (m, 6H), 1.43 (dt,J=8.6, 4.2 Hz, 1H), 1.38-1.21 (m, 3H), 1.18 (d, J=6.6 Hz, 3H), 1.15-1.05(m, 2H), 0.97-0.84 (m, 1H), 0.80 (t, J=7.3 Hz, 3H).

Example 12 Prophetic Synthesis of Compound 59

Compound 48: Compound 48 can be prepared in an analogous fashion tocompound 11, substituting 2, 2′-oxybis(ethylamine) for ethylenediamine.

Compound 59: Compound 59 can be prepared using an analogous sequence tocompound 24, substituting compound 48 for compound 11.

Example 13 Prophetic Synthesis of Compound 60

Compound 49: Compound 49 can be prepared in an analogous fashion tocompound 11, substituting butylenediamine for ethylenediamine.

Compound 60: Compound 60 can be prepared using an analogous sequence tocompound 24, substituting compound 49 for compound 11.

Example 14 Synthesis of Compound 30

Compound 27: Crotonic acid (1.15 g, 13.3 mmol) was dissolved in 30 mL ofwater. Sodium sulfite (2.03 g, 16 mmol) was added and the reactionmixture was stirred for 1 hour at 80° C. The reaction mixture was cooledon an ice bath and adjusted to pH=1 with concentrated HCl. The solventwas removed in vacuo to afford crude compound 27 which was used withoutfurther purification. MS negative mode: m/z=166.9 [M−H]⁻ C₄H₈O₅S(168.17).

¹H NMR (400 MHz, D₂O) δ 3.35 (s, 1H), 2.90 (dd, J=15.9, 5.3 Hz, 1H),2.47 (dd, J=15.9, 8.7 Hz, 1H), 1.33 (d, J=6.9 Hz, 3H).

Compound 30: To a solution of compound 27 (62 mg, 0.37 mmol) in waterwas added DIPEA (68 μL, 0.39 mmol), EDCI (75 mg, 0.39 mmol), and NHS (64mg, 0.56 mmol). The reaction mixture was stirred at room temperature for2 hours. Compound 11 (184 mg, 0.25 mmol) was added and the reactionmixture was stirred at room temperature overnight. The reaction mixturewas directly transferred to a 10 g C-18 cartridge and eluted with waterfollowed by 3/1 mater/methanol. Product containing fractions werecombined, concentrated, and lyophilized to afford 20 mg of compound 30as a white solid. MS negative mode: m/z=903.7 [M−Na]⁻, 881.7 [M−2Na]⁻,440.5 [(M−2Na)/2]⁻ C₃₈H₆₃N₃Na₂O₁₈S (927.36).

¹H NMR (400 MHz, Deuterium Oxide) δ 5.04 (d, J=4.0 Hz, 1H), 4.91 (q,J=6.4 Hz, 1H), 4.55 (d, J=8.8 Hz, 1H), 3.95 (dd, J=9.4, 3.6 Hz, 1H),3.89 (dd, J=14.6, 3.3 Hz, 2H), 3.80 (dd, J=10.8, 3.7 Hz, 2H), 3.76-3.70(m, 2H), 3.55 (t, J=6.0 Hz, 1H), 3.50 (t, J=6.2 Hz, 2H), 3.40-3.28 (m,3H), 3.13 (t, J=6.1 Hz, 2H), 2.44-2.31 (m, 1H), 2.23-2.11 (m, 1H), 2.06(d, J=4.4 Hz, 3H), 1.99-1.81 (m, 2H), 1.73 (q, J=13.9 Hz, 1H), 1.66-1.46(m, 2H), 1.45-1.31 (m, 1H), 1.29 (d, J=6.9 Hz, 1H), 1.24 (d, J=6.4 Hz,3H), 1.23-1.12 (m, 3H), 1.01-0.90 (m, 2H), 0.86 (t, J=7.3 Hz, 3H).

Example 15 Synthesis of Compound 31

Compound 28: Compound 28 was prepared in analogous fashion to compound27, starting with 4-bromocrotonic acid and using 2.4 equivalents ofsodium sulfite. MS negative mode: m/z=247.1 [M−H]⁻ C₄H₈O₈S₂ (247.97).

¹H NMR (400 MHz, Deuterium Oxide) δ 3.85-3.74 (m, 1H), 3.45 (dd, J=14.2,1.9 Hz, 1H), 3.03 (dd, J=14.3, 10.9 Hz, 1H), 2.96 (dd, J=16.6, 2.9 Hz,1H), 2.63 (dd, J=16.6, 9.4 Hz, 1H).

Compound 31: Compound 31 was prepared in an analogous fashion tocompound 30, from compound 11 and compound 28. MS negative mode:m/z=1007.5 [M-Na], 986.5 [M−2Na]⁻, 962.5 [M−3Na]⁻ C₃₈H₆₂N₃Na₃O₂₁S₂(1029.30).

¹H NMR (400 MHz, Deuterium Oxide) δ 5.03 (d, J=4.1 Hz, 1H), 4.92 (d,J=6.8 Hz, 1H), 4.56 (t, J=8.1 Hz, 1H), 4.03 (t, J=9.6 Hz, 1H), 3.90 (dd,J=10.6, 3.3 Hz, 1H), 3.86 (d, J=3.3 Hz, 1H), 3.80 (dd, J=8.9, 3.5 Hz,1H), 3.77 (d, J=4.0 Hz, 1H), 3.73 (d, J=6.0 Hz, 2H), 3.60 (t, J=5.9 Hz,1H), 3.48 (ddd, J=14.2, 5.3, 2.0 Hz, 2H), 3.43-3.36 (m, 1H), 3.33 (t,J=9.8 Hz, 2H), 3.30-3.21 (m, 1H), 3.11-3.05 (m, 1H), 3.05-2.99 (m, 1H),2.36 (t, J=12.5 Hz, 1H), 2.23 (d, J=11.8 Hz, 1H), 2.07 (s, 3H), 1.89 (d,J=12.7 Hz, 1H), 1.79 (m, 1H), 1.67 (d, J=20.5 Hz, 1H), 1.58 (d, J=16.5Hz, 1H), 1.53-1.42 (m, 1H), 1.34 (q, J=11.1, 10.2 Hz, 1H), 1.24 (d,J=6.5 Hz, 3H), 1.17 (q, J=12.4, 10.1 Hz, 2H), 1.02-0.91 (m, 1H), 0.87(t, J=7.5 Hz, 3H).

Example 16 Synthesis of Compound 32

Compound 29: Compound 29 was prepared in analogous fashion to compound27, starting with 4-bromocrotonic acid and using 1.05 equivalents ofsodium sulfite. MS negative mode: m/z=901.7 [M−Na]⁻, 879.7 [M−2Na]⁻,439.5 [(M−2Na)/2]⁻² C₃₈H₆₁N₃Na₂O₁₈S (925.35).

¹H NMR (400 MHz, Deuterium Oxide) δ 6.73 (dt, J=15.4, 7.6 Hz, 1H), 6.11(dd, J=15.6, 1.4 Hz, 1H), 3.81 (d, J=7.4 Hz, 2H).

Compound 32: Compound 32 was prepared in an analogous fashion tocompound 30, from compound 11 and compound 29. MS negative mode:m/z=896.3 [M−2Na]⁻ C₃₉H₆₅₁N₃Na₂O₁₈S (941.38).

¹H NMR (400 MHz, Deuterium Oxide) δ 6.69 (dt, J=15.3, 7.6 Hz, 1H), 6.19(d, J=15.4 Hz, 1H), 5.02 (d, J=4.0 Hz, 1H), 4.90 (q, J=8.5, 7.5 Hz, 1H),4.52 (d, J=8.6 Hz, 1H), 3.99 (t, J=9.6 Hz, 1H), 3.91-3.81 (m, 4H),3.81-3.75 (m, 3H), 3.72 (d, J=5.9 Hz, 3H), 3.56 (t, J=5.9 Hz, 1H),3.48-3.32 (m, 6H), 3.29 (q, J=7.1, 5.8 Hz, 4H), 3.13 (dq, J=22.3, 7.5Hz, 7H), 2.94 (s, 5H), 2.48 (s, 2H), 2.31 (t, J=12.7 Hz, 1H), 2.13 (d,J=5.4 Hz, 1H), 2.05 (s, 3H), 1.94 (p, J=6.0 Hz, 3H), 1.84 (d, J=11.5 Hz,1H), 1.78-1.66 (m, 1H), 1.66-1.42 (m, 3H), 1.41-1.28 (m, 1H), 1.23 (d,J=6.4 Hz, 3H), 1.15 (t, J=7.3 Hz, 7H), 1.06 (t, J=7.3 Hz, 1H), 0.93 (q,J=11.3 Hz, 1H), 0.84 (t, J=7.3 Hz, 3H).

Example 17 Synthesis of Compound 34

Compound 34: Compound 34 was prepared in an analogous fashion tocompound 30, from compound 11 and the commercially available5-sulfopentanoic acid. MS negative mode: m/z=896.3 [M−2Na]⁻C₃₉H₆₅₁N₃Na₂O₁₈S (941.38).

¹H NMR (400 MHz, D₂O) δ 4.93 (d, J=4.0 Hz, 1H), 4.81 (dd, J=8.0 Hz,J=12.0 Hz, 1H), 4.43 (broad d, 1H), 3.95-3.84 (m, 1H), 3.81-3.74 (m,2H), 3.73-3.68 (m, 2H), 3.67-3.58 (m, 4H), 3.50-3.44 (broad t, 1H),3.40-3.30 (m, 1H), 2.28-2.11 (m, 5H), 2.85-2.75 (m, 2H), 2.31-2.11 (m,J=8.0 Hz, 3H), 2.10-2.00 (m, 1H), 1.96 (s, 3H), 1.84-1.70 (m, 2H),1.69-1.34 (m, 11H), 1.33-1.18 (m, 3H), 1.17-0.94 (m, J=8.0 Hz, 8H),0.93-0.69 (m, J=8.0 Hz, 5H).

Example 18 Synthesis of Compound 33

Compound 33: Compound 32 (14 mg, 0.016 mmol) was dissolved in 1 mL ofwater. Three drops of saturated NaHCO₃ solution were added followed byPd/C (4 mg). The reaction mixture was vigorously stirred under ahydrogen atmosphere for 90 minutes. The reaction mixture was filteredthrough a 0.2 micron PTFE filter. The filtrate was applied to a 5 g C-18cartridge and eluted with water followed by 3/1 water/methanol. Productcontaining fractions were concentrated then lyophilized to afford 9 mgof compound 33 as a white solid (64%). MS negative mode: m/z=903.6[M-Na], 881.5 [M−2Na]⁻, 440.4 [(M−2Na)/2] C₃₈H₆₃N₃Na₂O₁₈S (927.36).

¹H NMR (400 MHz, Deuterium Oxide) δ 5.04 (d, J=4.0 Hz, 1H), 4.92 (q,J=6.4 Hz, 1H), 4.55 (d, J=8.6 Hz, 1H), 4.01 (t, J=9.5 Hz, 1H), 3.93-3.85(m, 3H), 3.85-3.77 (m, 2H), 3.73 (d, J=5.9 Hz, 2H), 3.62-3.55 (m, 1H),3.50-3.38 (m, 2H), 3.38-3.27 (m, 5H), 2.94-2.86 (m, 2H), 2.46-2.28 (m,4H), 2.16 (d, J=12.3 Hz, 1H), 2.07 (s, 3H), 2.04-1.96 (m, 2H), 1.86 (d,J=12.8 Hz, 2H), 1.72 (dd, J=38.2, 17.7 Hz, 1H), 1.63-1.44 (m, 3H),1.44-1.27 (m, 2H), 1.24 (d, J=6.5 Hz, 3H), 1.19-1.12 (m, 11H), 1.01-0.90(m, 1H), 0.86 (t, J=7.3 Hz, 3H).

Example 19 Synthesis of Compound 35

Compound 35: To a solution of compound 11 (29 mg, 39 μmole) in anhydrousDMSO (0.1 mL) was added a drop of DIPEA and the solution was stirred atroom temperature until a homogeneous solution was obtained. A solutionof succinic anhydride (4.1 mg, 41.2 μmole, 1.05 eq) in anhydrous DMSO(0.1 mL) was added in one portion and the resulting solution was stirredovernight. The solution was lyophilized to dryness and the crude productwas purified by HPLC to give compound 35 (28 mg, 34 μmole, 84%). MS:Calculated (C₃₈H₆₃N₃O₁₇, 833.4), ES-positive (856.3, M+Na), ES-negative(832.3, M−1).

¹H NMR (600 MHz, D₂O) δ 4.92 (d, J=4.0 Hz, 1H), 4.80 (dd, J=8.0 Hz, 1H),4.42 (d, J=8.0 Hz, 1H), 3.95-3.83 (m, 2H), 3.82-3.73 (m, 2H), 3.72-3.56(m, 5H), 3.49-3.41 (m, 1H), 3.40-3.31 (m, 1H), 3.30-3.11 (m, 5H), 2.55(t, J=8.0 Hz, 2H), 2.41 (t, J=8.0 Hz, 2H), 2.26-2.12 (m, 1H), 2.10-1.98(m, 1H), 1.95 (s, 3H), 1.84-1.68 (m, 2H), 1.67-1.34 (m, 8H), 1.33-1.17(m, 3H), 1.16-0.92 (m, J=8.0 Hz, 7H), 0.91-0.66 (m, J=8.0 Hz, 5H).

Example 20 Synthesis of Compound 36

Compound 36: Compound 36 was prepared in an analogous fashion tocompound 35, from compound 11 and glutaric anhydride. MS: Calculated(C₃₉H₆₅N₃O₁₇, 847.4), ES-positive (870.4, M+Na), ES-negative (846.3,M−1).

¹H NMR (400 MHz, D₂O) δ 4.98 (d, J=4.0 Hz, 1H), 4.86 (dd, J=8.0 Hz,J=12.0 Hz, 1H), 4.46 (broad d, 1H), 4.02 (dd, J=4.0 Hz, J=12.0 Hz, 1H),3.99-3.91 (m, 1H), 3.89 (m, 1H), 3.83 (dd, J=4.0 Hz, J=8.0 Hz, 1H),3.77-3.61 (m, 5H), 3.53-3.42 (m, 2H), 3.35-3.18 (m, 5H), 2.34 (t, J=8.0Hz, 2H), 2.31-2.18 (m, J=8.0 Hz, 3H), 2.13-2.04 (m, 1H), 2.01 (s, 3H),1.87-1.74 (m, J=4.0 Hz, 4H), 1.73-1.41 (m, 8H), 1.40-1.01 (m, J=8.0 Hz,10H), 0.96-0.75 (m, J=8.0 Hz, 5H).

Example 21 Prophetic Synthesis of Compound 61

Compound 50: Compound 50 can be prepared in an analogous fashion tocompound 11, substituting 1,4-xylylenediamine for ethylenediamine.

Compound 61: Compound 61 can be prepared in an analogous fashion tocompound 35, from compound 50 and succinic anhydride.

Example 22 Synthesis of Compound 43

Compound 39: To a solution of compound 38 (445 mg, 1.43 mmol) dissolvedin DMF (29 mL) at 0° C. was added HATU (563 mg, 1.14 mmol) and DIPEA(298 μL, 1.71 mmol). The mixture was stirred at 0° C. for 5 min. Asolution of compound 11 (840 mg, 1.14 mmol) in water (12 mL) was thenadded and the mixture was stirred for 12 h allowing to warm to roomtemperature. The volatiles were evaporated and the crude reactionmixture was co-evaporated 2× with toluene. The resulting yellow residuewas dried under high vacuum for 3 h. The residue was dissolved inpyridine (30 mL) at room temperature. Acetic anhydride (15 mL) was addedand the solution was stirred at r.t. for 12 h. The volatiles wereevaporated and the crude reaction mixture was co-evaporated 2× withtoluene. The resulting mixture was purified by column chromatography(DCM/MeOH 0→10%) to give compound 39 (1.197 g, 89%). LCMS (C-18; 5-95H₂O/MeCN): ELSD (peak at 4.804 min); UV (peak at 4.693 min).

Compound 40: A suspension of 39 (1.29 g, 1.096 mmol) and Pd/C 10% (50mg) in MeOH (45 mL) was degassed (3×) prior to the addition of hydrogenusing a balloon. The mixture was stirred at r.t. for 12 h. The catalystwas filtered off through a short pad of celite and the filtrate wasconcentrated under vacuum to give compound 40 as a white solid (1.13 g,99%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 3.998 min).

Compound 41: To a solution of compound 7 (140 mg, 0.345 mmol) dissolvedin DMF (14 mL) and cooled at 0° C. was added HATU (143 mg, 0.38 mmol)and DIPEA (76 μL, 0.435 mmol). The mixture was stirred at 0° C. for 5min. A solution of compound 40 (300 mg, 0.29 mmol) in DMF (4 mL) wasthen added and the mixture was stirred overnight allowing to warm toroom temperature. The volatiles were evaporated and the residueco-evaporated 2× with toluene. The resulting oil was purified by columnchromatography (DCM/MeOH 0→10%) to give compound 41 (317 mg, 76%). LCMS(C-18; 5-95 H₂O/MeCN): ELSD (peak at 5.352 min); UV (peak at 5.215 min).

Compound 42: A suspension of compound 41 (230 mg, 0.16 mmol) and Pd/C10% (10 mg) in MeOH (10 mL) was degassed (3×) prior to the addition ofhydrogen using a balloon. The mixture was stirred at r.t. for 16 h.Added more Pd/C 10% (10 mg) and stirred for additional 16 h. Reactionwas not complete. Added more Pd/C (10 mg) and stirred for another 4days. The catalyst was filtered off through a short pad of celite andthe filtrate concentrated under vacuum. The resulting residue waspurified by column chromatography (DCM/MeOH 0-30%) to give compound 42(123 mg, 67%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD (peak at 4.055 min).

Compound 43: A solution of sulfur trioxide pyridine (170 mg, 1.067 mmol)and compound 43 (124 mg, 0.106 mmol) in pyridine (4 mL) was stirred at67° C. for 30 min. The reaction mixture was concentrated under vacuum.The resulting yellow crude was dissolved in water and cooled to 0° C. A1N solution of NaOH was then added slowly until pH-10 and the mixturestirred overnight allowing to warm to room temperature. The aqueoussolution was lyophilized. The resulting white solid was dissolved inMeOH (5 mL), cooled to 0° C. and treated with NaOMe 25% solution in MeOH(0.5 mL). The mixture was stirred between 0° C. and 10° C. for 1 h. Thereaction mixture then neutralised by addition of 1N HCl until pH-7. TheMeOH was removed in vacuo. Water was added to the residue and thesolution was lyophilized. The resulting white solid was purified by C18ultra SNAP 30 g (3-5% MeOH/H₂O). Product containing fractions werecombined and lyophilized to afford compound 43 as an off-white solid(44.5 mg, 18% over 3 steps). LCMS: (C-18; 5-95 H₂O/MeCN): ELSD (peak at3.039 min), negative mode: m/z=584 [(M−2)/2]⁻ (loss of one sulfateduring ionisation), 624 [(M−2)/2]⁻. C₄₅H₇₈N₄O₃₀S₃ (1250) for compound 43and C₄₅H₇₈N₄O₂₇S₂ (1170) for disulfate.

¹H NMR (300 MHz, Deuterium Oxide) δ 5.01 (d, J=4.0 Hz, 1H), 4.91 (app d,J=6.2 Hz, 2H), 4.51 (d, J=8.6 Hz, 1H), 4.36-4.15 (m, 2H), 3.99 (t, J=9.5Hz, 1H), 3.92-3.68 (m, 7H), 3.68-3.54 (m, 5H), 3.49-3.37 (m, 3H),3.31-3.28 (m, 5H), 2.53 (t, J=5.4 Hz, 2H), 2.32 (t, J=12.5 Hz, 1H), 2.16(app d, J=12.3 Hz, 1H), 2.04 (s, 3H), 1.88-1.67 (m, 2H), 1.65-1.45 (m,3H), 1.39-1.25 (m, 1H), 1.22 (d, J=6.4 Hz, 3H), 1.18-1.12 (m, 1H),0.97-0.87 (m, 1H), 0.84 (t, J=7.4 Hz, 4H).

Example 23 Synthesis of Compound 44

Compound 44: Using the sequence of reactions in FIG. 12, compound 44 wasprepared substituting compound 8 for compound 7. LCMS: (C-18; 5-95H₂O/MeCN): ELSD (peak at 3.053 min), negative mode: m/z=584 [(M−2)/2]⁻(loss of one sulfate during ionisation), 624 [(M−2)/2]⁻. C₄₅H₇₈N₄O₃₀S₃(1250) for compound 44 and C₄₅H₇₈N₄O₂₇S₂ (1170) for disulfate.

¹H NMR (300 MHz, Deuterium Oxide) δ 5.07 (d, J=4.2 Hz, 2H), 4.96 (q,J=6.5 Hz, 1H), 4.93-4.83 (m, 1H), 4.57 (d, J=8.5 Hz, 1H), 4.31 (qd,J=10.8, 5.4 Hz, 2H), 4.05 (t, J=9.5 Hz, 1H), 3.95-3.88 (m, 2H),3.87-3.73 (m, 3H), 3.73-3.58 (m, 6H), 3.55-3.43 (m, 3H), 3.36 (s, 4H),2.58 (t, J=6.1 Hz, 2H), 2.38 (t, J=12.5 Hz, 1H), 2.22 (app d, J=12.3 Hz,1H), 2.09 (s, 3H), 1.90 (app d, J=12.0 Hz, 1H), 1.85-1.58 (m, 2H),1.58-1.32 (m, 1H), 1.28 (d, J=6.4 Hz, 3H), 1.24-1.16 (m, 1H), 1.05-0.94(m, 1H), 0.90 (t, J=7.3 Hz, 3H).

Example 24 Synthesis of Compound 46

Compound 45: To a stirred and cooled (15° C.) solution of propiolic acid(82 μl, 1.19 mmol) in DCM (5 mL) under argon was added oxalyl chloride(100 μl, 1.19 mmol) and DMF (5 μl, cat). The resulting mixture wasstirred for 20 min. A solution of compound 40 (180 mg, 0.17 mmol) inMeOH (20 mL) was treated with saturated sodium bicarbonate solution (4mL) 1 min prior to the addition of 3.5 ml of the propiolyl chloridesolution. The resulting mixture was stirred at room temperature for 40min. The reaction was filtered and partially concentrated under reducedpressure to the water fraction, which, was in-turn freeze-dried.Flash-column chromatography of the residue (0-10% MeOH in DCM) affordedcompound 45 as a white solid (110 mg, 60%). LCMS (C-18; 5-95 H₂O/MeCN):ELSD (peak at 3.38 min), UV (peak at 3.29 min), negative mode: m/z=1149[M+Na] C₅₃H₈₂N₄O₂₂ (1126).

Compound 46: A stirred solution of compound 45 (110 mg, 0.1 mmol) inMeOH (20 ml) under argon was treated with sodium methoxide until pH=9-10then stirred for 2 h. HPLC grade water (10 mL) was added and the mixtureconcentrated by rotary evaporation. The resulting aqueous solution waslyophilized. The residue was dissolved in HPLC grade water (20 ml) andthe pH adjusted to 9-10 with 1N NaOH and stirred at room temperature for2 h. The pH was adjusted to 8 (1M HCl) and the mixture lyophilized.Salts were removed by G10 gel permeation of the residue, (water aseluent).

The residue was combined with azide 3 in water (15 mL) under argon atroom temperature. A pre-mixed solution of copper sulfate and THPTA (0.43mL of 40 mmol aqueous solution) and sodium ascorbate (107 μmol, 0.107 mLof 1M solution) was added and the reaction mixture was stirred at roomtemperature for 2 hours. The reaction mixture was lyophilized. The crudereaction mixture was partially purified by P2 gel permeation (water aseluent) to give a yellow solid. This material was further purified byBiotage reverse phase C18 ultra (30 g) (3-5% H₂O/MeOH) chromatographyafforded compound 46 (54 mg, 46%). LCMS (C-18; 5-95 H₂O/MeCN): ELSD(peak at 3.08 min), UV (peak at 2.98 min), negative mode: m/z=671.2[(M−2)/2]⁻ C₄₉H₈₂N₇O₃₀S₃ (1344).

¹H NMR (300 MHz, Deuterium Oxide) δ 8.47 (s, 1H), 5.01 (d, J=4.0 Hz,1H), 4.90 (q, J=6.7 Hz, 1H), 4.71 (s, 2H), 4.51 (d, J=8.6 Hz, 1H), 4.05(s, 6H), 3.98 (t, J=9.4 Hz, 1H), 3.92-3.77 (m, 5H), 3.76-3.63 (m, 8H),3.57 (dt, J=11.3, 5.5 Hz, 3H), 3.43 (d, J=10.8 Hz, 1H), 3.36-3.19 (m,6H), 2.45 (t, J=6.0 Hz, 2H), 2.29 (app t, J=12.2 Hz, 1H), 2.15 (app d,J=11.8 Hz, 1H), 2.04 (s, 3H), 1.82 (app d, J=12.3 Hz, 2H), 1.78-1.42 (m,2H), 1.47-1.25 (m, 1H), 1.22 (d, J=6.5 Hz, 3H), 1.17-1.07 (m, 1H),0.97-0.88 (m, 1H), 0.82 (t, J=7.3 Hz, 3H).

Example 25 Prophetic Synthesis of Compound 73

Compound 63: A solution of compound 62 (preparation described in WO2007/028050) and phenyl acetylene (2.4 eq) in MeOH is degassed at roomtemperature. A solution of CuSO₄/THPTA in distilled water (0.04 M) (0.2eq) and sodium ascorbate (1.2 eq) is added successively and theresulting solution stirred 12 hrs at room temperature. The solution isconcentrated under reduced pressure. The crude product is purified byflash chromatography to give compound 63.

Compound 65: A mixture of compound 63 and compound 64 (preparationdescribed in WO 2013/096926) (1.7 eq) is azeotroped 3× from toluene. Themixture is dissolved in DCM under argon and cooled on an ice bath. Tothis solution is added boron trifluoride etherate (1.5 eq). The reactionmixture is stirred 12 hours at room temperature. The reaction isquenched by the addition of triethylamine (2 eq). The reaction mixtureis transferred to a separatory funnel and washed 1× half saturatedsodium bicarbonate solution and 1× water. The organic phase is driedover sodium sulfate, filtered, and concentrated. The residue is purifiedby flash chromatography to afford compound 65.

Compound 66: Compound 65 is dissolved in methanol at room temperature. Asolution of sodium methoxide in methanol (0.1 eq) is added and thereaction mixture stirred overnight at room temperature. The reactionmixture is quenched by the addition of acetic acid. The reaction mixtureis diluted with ethyl acetate, transferred to a separatory funnel andwashed 2× with water. The organic phase is dried over magnesium sulfate,filtered and concentrated. The residue is separated by flashchromatography to afford compound 66.

Compound 67: To a solution of compound 66 cooled on an ice bath is addedDABCO (1.5 eq) followed by monomethyoxytrityl chloride (1.2 eq). Thereaction mixture is stirred overnight allowing to warm to roomtemperature. The reaction mixture is concentrated and the residue ispurified by flash chromatography to afford compound 67.

Compound 68: To a solution of compound 67 in methanol is addeddibutyltin oxide (1.1 eq). The reaction mixture is refluxed for 3 hoursthen concentrated. The residue is suspended in DME. To this suspensionis added 2-(R)-[[(trifluoromethyl)sulfonyl]oxy]-cyclohexanepropanoicacid benzyl ester (preparation described in Thoma et. al. J. Med. Chem.,1999, 42, 4909) (1.5 eq) followed by cesium fluoride (1.2 eq). Thereaction mixture is stirred at room temperature overnight. The reactionmixture is diluted with ethyl acetate, transferred to a separatoryfunnel, and washed with water. The organic phase is dried over sodiumsulfate, filtered and concentrated. The residue is purified by flashchromatography to afford compound 68.

Compound 69: To a degassed solution of compound 68 in anhydrous DCM at0° C. is added Pd(PPh₃)₄ (0.1 eq), Bu₃SnH (1.1 eq) and acetic anhydride(2.0 eq). The resulting solution is stirred for 12 hrs while thetemperature is gradually increased to room temperature. The reactionmixture is diluted with DCM, transferred to a separatory funnel, andwashed with water. The organic phase is dried over Na₂SO₄, then filteredand concentrated. The residue is purified by flash chromatography toafford compound 69.

Compound 70: Compound 69 is dissolved in methanol and degassed. To thissolution is added Pd(OH)₂/C. The reaction mixture is vigorously stirredunder a hydrogen atmosphere for 12 hours. The reaction mixture isfiltered through a Celite pad and the cake is washed with MeOH. Thecombined filtrate is concentrated under reduced pressure. The crudeproduct is washed with hexane and dried under high vacuum to givecompound 70.

Compound 71: Compound 70 is dissolved in ethylenediamine and stirred at70° C. for 8 hours. The solvent is removed in vacuo and the crudematerial separated by C-18 reverse phase chromatography to affordcompound 71.

Compound 72: To a stirred and cooled (15° C.) solution of propiolic acidin DCM under argon is added oxalyl chloride and DMF (cat). The resultingmixture is stirred for 20 min. A solution of compound 72 in MeOHcontaining saturated sodium bicarbonate solution is added. The resultingmixture is stirred at room temperature for 30 min. The reaction isfiltered and concentrated under reduced pressure. Gel permeation of theresidue (water as eluent) affords compound 72.

Compound 73: A stirred solution of compound 72 and compound 3 in waterunder argon is treated with a pre-mixed solution of copper sulfate andTHPTA (40 mM aqueous solution) and sodium ascorbate. Upon completion thereaction is lyophilized. Gel permeation of the residue (water as eluent)affords compound 73.

Example 26 E-Selectin Activity—Binding Assay

The inhibition assay to screen for and characterize glycomimeticantagonists of E-selectin is a competitive binding assay, which allowsthe determination of IC₅₀ values. E-selectin/Ig chimera was immobilizedin 96 well microtiter plates by incubation at 37° C. for 2 hours. Toreduce nonspecific binding, bovine serum albumin was added to each welland incubated at room temperature for 2 hours. The plate was washed andserial dilutions of the test compounds were added to the wells in thepresence of conjugates of biotinylated, sLe^(a) polyacrylamide withstreptavidin/horseradish peroxidase and incubated for 2 hours at roomtemperature.

To determine the amount of sLe^(a) bound to immobilized E-selectin afterwashing, the peroxidase substrate, 3,3′,5,5′ tetramethylbenzidine (TMB)was added. After 3 minutes, the enzyme reaction was stopped by theaddition of H₃PO₄, and the absorbance of light at a wavelength of 450 nmwas determined. The concentration of test compound required to inhibitbinding by 50% was determined and reported as the IC₅₀ value for eachglycomimetic E-selectin antagonist as shown in the table below. IC₅₀values for exemplary compounds disclosed herein are provided in thefollowing table.

E-Selectin Antagonist Activity

Compound IC₅₀ (nM) 17 150 18 170 24 190 25 660 19 580 20 240 21 520 302777 31 2356 33 2115 34 340 36 510 35 720 43 106 44 136 46 192

1-53. (canceled)
 54. At least one compound chosen from glycomimeticE-selectin antagonists having the following Formulae:

and pharmaceutically acceptable salts thereof, wherein R¹ is chosen fromH, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₁₋₈ haloalkyl, C₂₋₈haloalkenyl, C₂₋₈ haloalkynyl,

groups, wherein n is chosen from integers ranging from 0 to 2, R⁷ ischosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆cycloalkylalkyl, and —C(═O)R⁸ groups, and each R⁸ is independentlychosen from H, C₁₋₈ alkyl, C₂₋₈ alkenyl, C₂₋₈ alkynyl, C₄₋₁₆cycloalkylalkyl, C₆₋₁₈ aryl, and C₁₋₁₃ heteroaryl groups; R³ is chosenfrom —OH, —OY¹, halo, —NH₂, —NHY¹, —NY¹Y², —OC(═O)Y¹, —NHC(═O)Y¹, and—NHC(═O)NHY¹ groups, wherein Y¹ and Y², which may be the same ordifferent, are independently chosen from C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl,C₂₋₁₂ alkynyl, C₄₋₁₆ cycloalkylalkyl, C₂₋₁₂ heterocyclyl, C₆₋₁₈ aryl,and C₁₋₁₃ heteroaryl groups, wherein Y¹ and Y² may join together alongwith the nitrogen atom to which they are attached to form a ring; and Mis chosen from linker groups.
 55. The at least one compound according toclaim 54, wherein the at least one compound is chosen from:


56. The at least one compound according to claim 54, wherein the atleast one compound is:


57. The at least one compound according to claim 56, wherein the atleast one compound is:


58. The at least one compound according to claim 56, wherein the atleast one compound is:


59. A composition comprising at least one compound according to claim 57and at least one additional pharmaceutically acceptable ingredient. 60.A method for decreasing the likelihood of the occurrence of; orabatement, lessening, decreasing occurrence, or alleviation of symptomsof; or delay or slowing of the progression of; or prolonging thesurvival of a subject afflicted with; or partial remission of; orameliorating at least one disease, disorder, and/or condition whereinhibition of E-selectin mediated functions is useful, the methodcomprising administering to a subject in need thereof an effectiveamount of at least one compound of claim
 57. 61. A method for decreasingthe likelihood of the occurrence of; or abatement, lessening, decreasingoccurrence, or alleviation of symptoms of; or delay or slowing of theprogression of; or prolonging the survival of a subject afflicted with;or partial remission of; or ameliorating at least one inflammatorydisease, disorder, and/or condition, the method comprising administeringto a subject in need thereof an effective amount of at least onecompound of claim
 57. 62. A method for decreasing the likelihood of theoccurrence of; or abatement, lessening, decreasing occurrence, oralleviation of symptoms of; or delay or slowing of the progression of;or prolonging the survival of a subject afflicted with; or partialremission of; or ameliorating cancer, the method comprisingadministering to a subject in need thereof an effective amount of atleast one compound of claim
 57. 63. The method according to claim 62,wherein the cancer is chosen from solid tumor cancers.
 64. The methodaccording to claim 62, wherein the cancer is chosen from bone cancers,colorectal cancers, and pancreatic cancers.
 65. The method according toclaim 62, wherein the cancer is chosen from liquid tumor cancers. 66.The method according to claim 62, wherein the cancer is chosen fromacute myelogenous leukemia, acute lymphoblastic leukemia, chronicmyelogenous leukemia, and multiple myeloma.
 67. A method for decreasingthe likelihood of the occurrence of; or abatement, lessening, decreasingoccurrence, or alleviation of symptoms of; or delay or slowing of theprogression of; or prolonging the survival of a subject afflicted with;or partial remission of; or ameliorating cancer, the method comprisingadministering to a subject in need thereof (a) an effective amount of atleast one compound of claim 57 and (b) at least one of therapy chosenfrom (i) chemotherapy and (ii) radiotherapy.
 68. A method for decreasingthe likelihood of the occurrence of; or abatement, lessening, decreasingoccurrence, or alleviation of symptoms of; or delay or slowing of theprogression of; or prolonging the survival of a subject afflicted with;or partial remission of; or ameliorating metastasis of cancer cells, themethod comprising administering to a subject in need thereof aneffective amount of at least one compound of claim
 57. 69. A method forinhibiting infiltration of cancer cells into the liver, lymph nodes,lung, bone, and/or bone marrow, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofclaim
 57. 70. A method for enhancing hematopoietic stem cell survival,the method comprising administering to a subject in need thereof aneffective amount of at least one compound of claim
 57. 71. The methodaccording to claim 70, wherein the subject has cancer and has receivedor will receive chemotherapy and/or radiotherapy.
 72. A compositioncomprising at least one compound according to claim 58 and at least oneadditional pharmaceutically acceptable ingredient.
 73. A method fordecreasing the likelihood of the occurrence of; or abatement, lessening,decreasing occurrence, or alleviation of symptoms of; or delay orslowing of the progression of; or prolonging the survival of a subjectafflicted with; or partial remission of; or ameliorating at least onedisease, disorder, and/or condition where inhibition of E-selectinmediated functions is useful, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofclaim
 58. 74. A method for decreasing the likelihood of the occurrenceof; or abatement, lessening, decreasing occurrence, or alleviation ofsymptoms of; or delay or slowing of the progression of; or prolongingthe survival of a subject afflicted with; or partial remission of; orameliorating at least one inflammatory disease, disorder, and/orcondition, the method comprising administering to a subject in needthereof an effective amount of at least one compound of claim
 58. 75. Amethod for decreasing the likelihood of the occurrence of; or abatement,lessening, decreasing occurrence, or alleviation of symptoms of; ordelay or slowing of the progression of; or prolonging the survival of asubject afflicted with; or partial remission of; or ameliorating cancer,the method comprising administering to a subject in need thereof aneffective amount of at least one compound of claim
 58. 76. The methodaccording to claim 75, wherein the cancer is chosen from solid tumorcancers.
 77. The method according to claim 75, wherein the cancer ischosen from bone cancers, colorectal cancers, and pancreatic cancers.78. The method according to claim 75, wherein the cancer is chosen fromliquid tumor cancers.
 79. The method according to claim 75, wherein thecancer is chosen from acute myelogenous leukemia, acute lymphoblasticleukemia, chronic myelogenous leukemia, and multiple myeloma.
 80. Amethod for decreasing the likelihood of the occurrence of; or abatement,lessening, decreasing occurrence, or alleviation of symptoms of; ordelay or slowing of the progression of; or prolonging the survival of asubject afflicted with; or partial remission of; or ameliorating cancer,the method comprising administering to a subject in need thereof (a) aneffective amount of at least one compound of claim 58 and (b) at leastone of therapy chosen from (i) chemotherapy and (ii) radiotherapy.
 81. Amethod for decreasing the likelihood of the occurrence of; or abatement,lessening, decreasing occurrence, or alleviation of symptoms of; ordelay or slowing of the progression of; or prolonging the survival of asubject afflicted with; or partial remission of; or amelioratingmetastasis of cancer cells, the method comprising administering to asubject in need thereof an effective amount of at least one compound ofclaim
 58. 82. A method for inhibiting infiltration of cancer cells intothe liver, lymph nodes, lung, bone, and/or bone marrow, the methodcomprising administering to a subject in need thereof an effectiveamount of at least one compound of claim
 58. 83. A method for enhancinghematopoietic stem cell survival, the method comprising administering toa subject in need thereof an effective amount of at least one compoundof claim
 58. 84. The method according to claim 83, wherein the subjecthas cancer and has received or will receive chemotherapy and/orradiotherapy.